Oxygenated polymethyleneimines

ABSTRACT

WHEREIN N3 IS A WHOLE NUMBER FROM 1 TO 3, INCLUSIVE, N4 IS A WHOLE NUMBER FROM 3 TO 5. INCLUSIVE, AND N5 IS A WHOLE NUMBER FROM 6 TO 8, INCLUSIVE, IN WHICH THE SUM OF N3 AND N4 IS NOT LESS THAN 5 AND NOT GREATER THAN 7 AND THE SUM OF N4 AND N5 IS 11; W IS SELECTED FROM THE GROUP CONSISTING OF CARBONYL, HYDROXYMETHYLENE, ACYLOXYMETHYLENE, METHYLHYDROXYMETHYLENE, PHENYLHYDROXYMETHYLENE, ALKYLENEDIOXYMETHYLENE, AND CYCLICAMINOMETHYLENE; W1 IS SELECTED FROM THE GROUP CONSISTING OF CARBONYL, HYDROXYMETHYLENE, ACYLOXYMETHYLENE AND ALKYLENEDIOXYMETHYLENE; Z1 IS SELECTED FROM THE GROUP CONSISTING OF HYDROGEN, CYCLOALKYL, CYANOMETHYL, 2-AMINOETHYL, 2-GUANIDINOETHYL, OXYGENATED POLYMETHYLENEIMINES OF THE FORMULAE   Z1-N&lt;(-(CH2)N3-W-(CH2)N4-), OR Z-N&lt;(-(CH2)N4-W1-(CH2)N5-)     -CO-R, -CH2-R, -SO2-R, AND -COO-R1   IN WHICH R IS ARYL AND R1 IS ARALKYL, AND THE PHARMACOLOGICALLY ACCEPTABLE ACID ADDITION SALTS OF THOSE COMPOUNDS WHEREIN THE GROUP, &gt;N-Z1 IS AN AMINO RADICAL; AND Z IS SELECTED FROM THE GROUP CONSISTING OF   -CO-R, -SO2-R, AND -COO-R1   IN WHICH R AND R1 HAVE THE MEANINGS GIVEN ABOVE. THE COMPOUNDS OF THE ABOVE FORMULAE AND THEIR ACID ADDITION SALTS ARE PARTICULARLY VALUABLE AS CENTRAL NERVOUS SYSTEM STIMULANTS.

nitedStates Patent Ofice 3,780,022 Patented Dec. 18, 1973 ABSTRACT OFTHE DISCLOSURE Oxygenated polymethyleneimines of the formulae wherein n,is a whole number from 1 to 3, inclusive, n is a whole number from 3 to5, inclusive, and n is a whole number from 6 to 8, inclusive, in whichthe sum of n;, and n is not less than 5 and not greater than 7 and thesum of 11 and n is 11; W is selected from the group consisting ofcarbonyl, hydroxymethylene, acyloxymethylene, methylhydroxymethylene,phenylhydroxymethylene, alkylenedioxymethylene, andcyclicaminomethylene; W is selected from the group consisting ofcarbonyl, hydroxymethylene, acyloxymethylene and alkylenedioxymethylene;Z is selected from the group consisting of hydrogen, cycloalkyl,cyanomethyl, 2-aminoethyl, 2-guanidinoethyl,

.O O R, CHa-R, --S Oz-R and ii-OR in which R is aryl and R is aralkyl,and the pharmacologically acceptable acid addition salts of thosecompounds wherein the group, N-Z is an amino radical; and Z is selectedfrom the group consisting of in which R and R have the meanings givenabove. The compounds of the above formulae and their acid addition saltsare particularly valuable as central nervous system stimulants.

CROSS REFERENCE This application is a continuation-in-part ofapplication Ser. No. 666,991 filed Sept. 11, 1967, now US. Pat. 3,556,943. Application Ser. No. 666,991 is a continuationin-part ofabandoned application Ser. No. 453,204, filed May 4, 1965.

BACKGROUND OF THE INVENTION This invention relates to a bioconversionprocess to obtain oxygenated polymethyleneimines and azabicycloalkanes,some of which are known but which have not hitherto been readilyavailable by purely chemical methods of synthesis.

SUMMARY OF THE INVENTION This invention relates to a novel method forthe introduction of oxygen into the heterocyclic ring of N-acylderivatives of polymethyleneimines (azacycloalkanes) andazabicycloalkanes. More particularly this invention relates to theintroduction of oxygen into the heterocyclic ring of N-aromatic acylderivatives of polymethyleneimines (azacycloalkanes) andazabicycloalkanes by subjecting them to the oxygenating activity of themicroorganism Sporotrichum sulfurescens, to certain novel productsproduced by the process of this invention and to derivatives thereof.

DETAILED DESCRIPTION OF THE INVENTION The process of this invention isof particular value for the introduction of oxygen into theheterocyclic'ring of compounds having the formula:

L X1 I in which R is aryl, and R is aralkyl.

The above compounds of Formula I when subjected to the bioconversionprocess of this invention give rise to the corresponding compoundswherein the heterocyclic ring has been oxygenated by the introduction ofa hydroxy or keto group as represented by Formula Ia.

wherein X, X, and Z have the meanings previously given and Y is hydroxyor keto.

The bioconversion process of this invention when applied to thecompounds of Formula I, which contain one or more attached alkyl groups,namely, the compounds of Formula I wherein X and X are methyl, ethyl orpropyl also give rise to the corresponding compounds wherein the alkylside chain has been oxygenated by the introduction of a hydroxy or ketogroup. These compounds are likewise useful for the same purposes ashereinafter given for the compounds in which the heterocyclic ring hasbeen oxygenated.

The bioconversion process of this invention is of particular value forthe production of compounds having the formula:

X 1) ul wherein X, X and Z have the meanings previously given, n is awhole number from 1 to 5, inclusive, and n is a whole number from 2 to8, inclusive, in which the sum of In and n is not less than 3 and notmore than 11.

The compounds produced by the bioconversion process" the correspondingketo compounds, particularly those having the formula:

X l)nl O=O N-Z (CW2): X1

III

wherein n n X, X and Z have the meanings and limitations previouslygiven.

The compounds produced by the bioconversion process wherein theintroduced oxygen substituent is keto, or the keto compounds produced bythe above chemical oxi dation step, can if desired be reduced by methodshereinafter disclosed, for example using sodium borohydride, to obtainthe corresponding hydroxy compounds.

While the process of the invention is of general applicability and canbe employed for the oxygenation of all of the N-acyl derivatives ofpolymethyleneimines and azabicycloalkanes of Formula I, above, to obtainthe oxygenated compounds of Formulae II and III, above, the process ofthe invention is especially advantageous for obtaining compounds of theinvention as represented by Formulae IV, V, VI and VII below:

VII

wherein n is a whole number from 1 to 3, inclusive, n is a whole numberfrom 3 to 5, inclusive, and n is a whole number from 6 to 8, inclusive,in which the sum of n and n is not less than 5 and not greater than 7and the sum of n; and n is 11, n is 0 or 1 and n is 0 or 1, in which 11and n cannot each equal 0 simultaneously; W is a radical selected fromthe group consisting of carbonyl, hydroxymethylene, acyloxymethylene,methylhydroxymethylene, phenylhydroxymethylene, alkylenedioxymethylene,and cyclicaminomethylene; Z is selected from the group consisting ofhydrogen, cycloalkyl, cyanomethyl, Z-aminoethyl, Z-guanidinoethyl,

in which R is aryl and R is aralkyl, and Y is hydrogen, halogen, hydroxyor alkoxy.

In this application the term aryl means an aryl radical of 6 to 12carbon atoms, inclusive, such as, for example, phenyl, tolyl, xylyl,naphthyl, biphenylyl and the like. The term aralkyl means an aralkylradical of 7 to 16 carbon atoms, inclusive, such as benzyl,p-nitrobenzyl, 4-methylbenzyl, 3-methylbenzyl, 4-methylphenethyl,4-biphenylbutyl, a-naphthylmethyl, B-naphthylethyl, and the like. Theterm acyloxymethylene means a substituent wherein the acyl radical isthat of an organic carboxylic acid of from 1 to 16 carbon atoms,inclusive,

is a saturated 5 to 9 ring atom cyclic amino radical such as thosehereinafter listed. The term cycloalkyl means a cycloalkyl radical of 5to 15 carbon atoms inclusive, such as cyclopentyl, cyclohexyl,cyclooctyl, cyclodecyl, cyclopentadecyl and the like. The term alkoxymeans an alkoxy radical wherein alkyl is an alkyl radical of 1 to 6carbon atoms.

The oxygenated compounds produced by the bioconin which the symbolversion and oxidation processes of this invention and the derivativesthereof, including those represented by Formulae II, III, IV, V, VI andVII, above; and the compounds produced in the Examples appended heretoare useful as insecticides, fungicides, parasiticides, proteindenaturants, insect repellants, high boiling solvents, plasticizers forsynthetic resins, crosslinking agents for fiber synthesis, pharmacologicagents for psychic control effects and as intermediates for dyes,polymers and fibers.

The compounds of Formulae IV, V, VI and VII, above, and thepharmacologically acceptable acid addition salts of those compoundswhich contain an amino radical are of particular value as centralnervious system stimulants, they are mood elevator and psychicenergizers which are useful in the treatment of mental healthconditions. In addition the compounds of Formula VI wherein Z is arefungicidal agents which are useful for the treatment of fungalinfections. For example, benzyl6-oxo-3-azabicyclo[3.2.2]nonane-B-carboxylate is active againstMonilinia fructicola.

As central nervious system stimulants and fungicidal agents thecompounds of Formulae IV, V, VI and VII of this invention can beprepared and administered to humans, mammals, birds and animals, in awide variety of oral or parenteral dosage forms, singly or in admixturewith other coacting compounds, in doses of from about 0.1 to aboutmg./kg., depending on the severity of the condition being treated andthe recipients response to the medication. They can be administered witha pharmaceutical carrier which can be a solid material or a liquid inwhich the compound is dissolved, dispersed or suspended. The solidcompositions can take the form of tablets, powders, capsules, pills, orthe like, preferably in unit dosage forms for simple administration orprecise dosages. The liquid compositions can take the form of solutions,emulsions, suspensions, syrups or elixirs.

As an example of their use as intermediates the compounds wherein theoxygen substituent is keto (the hydroxy compounds can be oxidized toketo) can be converted to lactams which can be hydrolyzed to amino acidsin accordance with the procedures disclosed in US. Pats. 2,579,851 and2,569,114. For example, the ketones are converted to the oximes byreacting them with hydroxylamine or a salt thereof. The oximes are thensubjected to a Beckman rearrangement by treatment with sulfuric acid orthe equivalent to produce lactams. The lactams thus produced are usefulintermediates giving on hydrolysis amino acids. The lactams and aminoacids thus obtained are useful for the manufacture of valuable products,for example, polyamides, as disclosed in U.S. Pat. 2,579,851, supra.

For use as insect repellents the compounds of this invention can beformulated with aqueous or nonaquous carriers in accordance with methodsknown in the art, for example, U.S. Pat. 3,131,215.

The microbiological process of this invention comprises subjecting anN-aromatic acyl derivative of a methyleneimine (I) to the oxygenatingactivity of the species of fungus Sporotrichum sulfurescens. The genusS'porotrichum belongs to the family Moniliaceae of the order Monilialesof the class Deuteromycetes.

The typical strain preferred for the practice of this invention isSporotrichum sulfurescens, available from the American Type CultureCollection, Washington, D.C., Collection No. ATCC 7159. It is to beunderstood, however, that other strains of Spamtrichum sulfurescens aresuitable for the practice of this invention.

The starting materials (I) for the process of this invention, some ofwhch are known, are prepared from polymethyleneimines (azacycloalkanes)and azabcycloalkanes of the formula:

wheren n, X and X have the meanings previously given, by methods knownin the art for acylating polymethyleneimines. For example, the selectedpolymethyleneimine or azabicycloalkane is dissolved, mixed or suspendedin an aqueous sodium hydroxide solution and reacted with the acid halideof the appropriate monobasic aryl or aralkyl carboxylic acid, such asthose, hereinafter listed as acylating agents, or with the acid halideof the appropriate monobasic aryl sulfonic acid such, as for example,benzenesulfonic acid, m-, and p-toluenesulfonic acids, aandi-naphthalene sulfonic acids, p-chlorobenzenesulfonic acid and the like,as illustrated by preparations 1 through 10, herein. The acylation mayalso be carried out in hydrocarbon solvents such as, e.g., Skellysolve B(mixed hexanes), benzene or toluene. In carrying out the bioconversionprocess of this invention the operational conditions and reactionprocedures are advantageously those known in the art of bioconversion asillustrated in Murray et al., U.S. Pats. 2,602,769 and 2,735,800,utilizing the oxygenating activity of the microorganism Sporatrichumsulfurescens.

In the practice of this invention, the bioconversion can be effected bya growing or resting culture of the microorganism or by spores, washedcells or enzymes of the microorganism.

Culture of the microorganism for the purpose and practice of thisinvention is in or on a medium favorable to development of themicroorganism. Sources of nitrogen and carbon should be present in theculture medium and an adequate sterile air supply should be maintainedduring the conversion, for example, by the conventional techniques ofexposing a large surface of the medium or by passing air through asubmerged culture.

Nitrogen in assimilable form can be provided by sources normallyemployed in such processes, such as corn steep liquor, soybean meal,yeast extracts, peptone, soluble or insoluble vegetable or animalprotein, lactalbumin, casein, whey, distillers solubles, amino acids,ni-

6 trates and ammonium compounds, such as ammonium tartrate, nitrate,sulfate and the like.

Available carbon can also be provided by sources normally used inbioconversions such as carbohydrates, e.g., glucose, fructose, sucrose,lactose, maltose, dextrines, starches; meat extracts, peptones, aminoacids, proteins, fatty acids, glycerol, whey and the like. Thesematerials may be used either in a purified state or as concentrates suchas whey concentrate, corn steep liquor, grain mashes, and the like, oras mixtures of the above. Many of the above sources of carbon can alsoserve as a source of nitrogen.

The medium can desirably have a pH before inoculation of between about 4to about 7 though a higher or lower pH can be used. A temperaturebetween about 25 to 32 C. is preferred for growth of the microorganismbut higher or lower temperatures within a relatively wide range aresuitable.

The substrate can be added to the culture during the growth period ofthe microorganism as a single feed or by gradual addition during theconversion period or it can be added to the medium before or aftersterilization or inoculation making appropriate adjustments for effectsof pH and/or temperature upon the stability of the substrate used. Thepreferred, but not limiting, range of concentration of the substrate inthe culture medium is about 0.1 to 10 grams per liter. The substrate isadded to the medium in any suitable manner, especially one whichpromotes a large surface contact of the substrate to the oxidizingactivity of the microorganism, for example, by dissolving the substrate,when it is a solid, in an organic solvent and mixing the solutionthoroughly with the medium or by adding to the medium finely comminutedparticles of the substrate, e.g., micronized particles, preferably byweight smaller than 20- microns, either as a dry powder or, preferablyfor mechanical reasons, as an aqueous suspension. In preparing theaqueous suspension, the use of dispersing or suspending agent isadvantageous.

The temperature during the fermentation can be the same as that foundsuitable for growth of the microorganism. It need be maintained onlywithin such range as supports life, active growth or the enzyme activityof the microorganism; the range of 20 to 35 C. is preferred. A pH ofabout 4 to 6 is generally preferred for growth of the microorganismduring the bioconversion but for acid-sensitive substrates, the pHshould be about 7 during the fermentation. Aeration can be effected bysurface culture or preferably under submerged fermentation conditions,in accordance with methods well known in the art. The time required foroxygenation by the enzymatic system of the microorganism employed canvary considerably. The range of about 2 to hours is practical but notlimiting; 72 hours is generally satisfactory. The progress of thebioconversion and its completion are conveniently determined bypaper-strip chromatography, vapor-phase chromatography or thin-filmchromatography [Heftman, Chromatography (1961) Reinhold Publishing Co.,New York, N.Y.].

Alternatively, the oxygenation of the selected substrate can be effectedunder aerobic conditions by subjecting it to the oxygenating action ofoxygenating enzymes isolated from the microorganism, to the action ofspores of the microorganism, and to the action of isolated cells of themicroorganism. Isolated enzyme preparations can be prepared inaccordance with the general procedure disclosed by Zuidweg et al.,Biochim. Biophys. Acta, 58, 131-133 (1962). Oxygenation can be effectedwith spores in accordance with the general process disclosed in U.S.Pats. 3,031,379 and 3,031,382. The separation of washed cells from thefermentation medium is well known in the art, see for example U.S. Pat.2,831,789.

The term oxygenating activity as used throughout this specificationmeans the enzymatic action of a growing or resting culture of themicroorganism or of spores, Washed cells or isolated enzymes of themicroorganism,

7 which elfects introduction of oxygen in the molecule of the substrate,under aerobic fermentation conditions.

After completion of the fermentation, the resulting oxygenated productsare recovered from the fermentation beer by conventional methods. Forexample, the whole beer can be extracted with a water-miscible organicsolvent such as methylene chloride, chloroform, carbon tetrachloride,ethylene chloride, trichloroethylene, ether, amyl acetate, benzene, andthe like or the beer and mycelia can be separated by conventionalmethods such as centrifugation or filtration, and then separatelyextracted with suitable solvents. The mycelia can be extracted witheither water-miscible or water-immiscible solvents or in cases wherelittle or no product is contained in the mycelium, it can be merelywashed with water and the wash water added to the beer filtrate. Thebeer, free of mycelia, can then be extracted with water-immisciblesolvents such as those listed above. The extracts are combined, driedover a drying agent such as anhydride sodium sulfate, and the solventremoved by conventional methods such as evaporation or distillation atatmospheric or reduced pressure. The oxygenated products thus obtainedcan be further purified by conventional methods, e.g.,recrystallization, chromatography, distillation in the case of liquids,and the like.

Separation of the various oxygenated products obtained from thefermentation can be accomplished by conventional methods such aschromatography and/ or fractional crystallization and, if liquids, bydistillation. In instances when separation of the hydroxy compounds isdifiicult a convenient and advantageous method is first to oxidize underacidic neutral or slightly basic conditions the crude oxygenatedpolymethyleneimines or azabicycloalkanes obtained from the beer inaccordance with methods known in the art for oxidizing secondary hydroxygroups to ketones, for example, Fieser and Fieser, Natural ProductsRelated to Phenanthrene, 3rd ed., pages 127-129, 193 and 194, ReinholdPublishing Corporation, New York, N.Y. Thus, the crude bioconversionproducts are dissolved in an inert organic solvent such as acetone,benzene, methylene chloride, t-butanol, and the like, and then oxidizedwith aqueous chromic acid, potassium permanganate, tbutyl hypochloriteor like oxidizing agents to convert the secondary hydroxy groups presentto keto, thereby producing a mixture of the corresponding keto compoundswhich are in some cases more easily separated by chromatography and/orcrystallization or distillation in the case of liquids.

The compounds obtained from the bioconversion wherein the introducedoxygen substituent is keto or those prepared by the above chemicaloxidation of the corresponding hydroxy compounds, for example, compoundsof Formula III, can, if desired, be reduced, preferably under neutral oracidic conditions, in accordance with methods known in the art forreducing carbonyl groups to produce the corresponding hydroxy compounds.For example, reduction can be conveniently accomplished with hydrogen inthe presence of a catalyst such as palladium, platinum or Raney nickelunder neutral conditions; sodium in an alkanol; or with a reducing agentsuch as lithium aluminum hydride, sodium borohydride, primary isobutylmagnesium bromide or lithium tertiary butoxy aluminum hydride, and thelike.

The compounds obtained from the bioconversion wherein the introducedoxygen substituent is hydroxy, for example the compounds of Formula II,can be acylated to give the corresponding acyloxy compounds inaccordance with methods known in the art for acylating secondary hydroxygroups, for example, by reaction with the appropriate acid anhydride oracid halide, by reaction with the appropiate ester or by reaction withthe appropriate acid in the presence of an esterificatiou catalyst, etc.Suitable acylating agents are organic carboxylic acids, particularlyhydrocarbon carboxylic acids containing from 1 to 16 carbon atoms,inclusive, or acid anhydrides or acid halides thereof. Illustrative ofhydrocarbon carboxylic acids employed in the formation of the acylatesare saturated and unsaturated aliphatic acids and aromatic acids such asacetic, propionic, butyric, isobutyric, tert.-butylacetic, valeric,isovaleric, caproic, caprylic, decanoic, dodecanoic, acrylic, crotonic,hexynoic, heptynoic, octynoic, cyclobutanecarboxylic,cyclopentanecarboxylic, cyclopentenecarboxylic, cyclohexanecarboxylic,dimethylcyclohexanecarboxylic, benzoic, toluic, naphtholic,ethylbenzoic, phenylacetic, naphthaleneacetic, phenylvaleric, cinnamic,phenylpropiolic, phenylpropionic, p-butoxyphenylpropionic, succinic,glutaric, dimethylglutaric, maleic, cyclopentylpropionic acids, and thelike. If the acylating agent is free acid, the reaction is preferablyeffected in the presence of an esterification catalyst, for examplep-toluenesulfonyl chloride, trifiuoroacetic anhydride, p-toluenesulfonicacid, trifluoroacetic acid, sulfuric acid, and the like.

The compounds obtained from the bioconversion process wherein theintroduced oxygen substituent is keto or the keto compounds obtained bychemical oxidation of the corresponding hydroxy compounds such as thecompounds of Formula III, can be converted to their common carbonylderivatives such as oximes, hydrazones, semicarbazones, cyclic alkyleneketals and the like in accordance with methods well known in the art.For example, the carbonyl group can be ketalized by reacting theselected compound with an alkanediol selected from the group of vicinalalkane-1,2-diols and alkane-1,3-diols containing up to and including 8carbon atoms, e.g., ethylene, propylene, trimethylene, 2,3 butylene,2,4-pentylene, 4-methyl-1,2-pentylene, 1,3-hexylene, 1,2-heptylene, 3,4-heptylene, 1,3-octylene, and the like, preferably in an organic solventsuch as benzene, toluene, xylene, methylene chloride, and the like andin the presence of an acid catalyst such as p-toluenesulfonic' acid. Thereaction is conducted at a temperature between about 20 and about 2000., preferably between about 40 and about C. The time required for thereaction is not critical and may be varied between about 1 and 48 hours,depending on the temperature.

The acylates and the carbonyl derivatives can, if desired, be removed byhydroysis in accordance with methods known in the art, e.g., with diluteacids or bases.

Other derivatives of the oxygenated bioconversion products of theinvention can be prepared in accordance with methods known in the art;for example, the compounds which have the radical,

attached to the nitrogen can be reduced using lithium aluminum hydrideor diborane to obtain the corresponding aralkyl amines having thepartial formula,

If additional unprotected carbonyl groups are present in the moleculethey will be concomitantly reduced to hydroxymethylene, and can beoxidized back to carbonyl, if desired, using chromic acid (Jonesreagent) as hereinbefore described. The aralkyl amines thus obtained canbe hydrogenolyzed, eg using hydrogen in the presence of a catalyst, suchas palladium on carbon or Raney nickel to give the correspondingsecondary amine, NH. The secondary amine, thus obtained can be reactedwith chloroacetonitrile in the presence of sodium carbonate to obtainthe corresponding cyanomethylarnine, which is reduced with lithiumaluminum hydride to the corresponding 2-aminoethyl derivative having thepartial formulas, NCH CH NH The aminoethyl compound thus ob tained isthen reacted with 2-methyl-2-thiopseudourea sulfate to obtain thecorresponding Z-guanidinoethylamine sulfate having the partial formula:

which can be neutralized with alkali to obtain the corresponding freebase. The secondary amines obtained, above, can be cycloalkylated at thenitrogen group by reacting with a cycloalkanone followed by reductionwith lithium aluminum hydride to obtain the corresponding cycloalkylamines.

The compounds of the invention wherein the substituent Z or Z is whereinR and R have the meaning previously given, and which have a hydroxygroup attached to the heterocyclic ring, can be subjected to acylmigration, e.g. with hydrochloric acid in tetrahydrofuran, which resultsin acylation of the hydroxy group to obtain the corresponding secondaryamine.

The compounds of this invention, wherein the substituent Z or Z is:

in which R is aryl and R is aralkyl and which have a keto group attachedto the heterocyclic ring, can be halogenated on the heterocyclic ringand then subjected to solvolysis with an alcohol in the presence ofsodium hydroxide to obtain the corresponding alkyloxy compound. Thecompounds of this invention having the substituents described above canalso be reacted with a Grignard reagent, RMgX, wherein R is alkyl oraryl, as hereinbefore defined and X is halogen, in accordance with knownmethods to obtain the corresponding compounds wherein the ketosubstituent is replaced by hdroxy and the R radical of the particularGrignard reagent employed. The acyl substituent on the nitrogen isreplaced by hydrogen during the Grignard reaction.

The following preparations and examples are intended to illustrate theprocess as applied to representative and typical starting materials. Thefollowing examples are for the purpose of illustrating the best modecontemplated of carrying out the invention and to supplement theforegoing disclosure of the invention with additional descriptions ofthe manner and process of carrying out the invention so as further toenable workers skilled in the art to do so.

In addition to the uses given hereinabove, the compounds of thisinvention which are free amines form salts with fluosilicic acid whichare useful as mothproofing agents in accordance with US. Pats. 1,915,334and 2,075,- 359. These amines also form salts with thiocyanic acid whichcondense with formaldehyde to form resinous materials useful as picklinginhibitors according to US. Pats. 2,425,320 and 2,606,155.

PREPARATION 1.1-BENZOYLHEXA- METHYLENEIMINE 'Benzoyl chloride (60 ml.)in 200 ml. of Skellysolve B was added to a stirred, cooled (ice-bath)solution of 200 ml. of hexamethyleneimine in 800 ml. of Skellysolve B.The mixture was then washed several times with 1 N hydrochloric acid andwith water, and filtered through anhydrous sodium sulfate. Evaporationof the Skellysolve B hexanes (hereinafter called Skellysolve B) anddistillation of the oily residue gave 40.5 g. ofl-benzoylhexamethyleneimine, -B.P. 156-160/ 1 torr.

Analysis.-Calcd. for C H NO (percent): C, 76.81; H, 8.43; N, 6.89. Found(percent): C, 75.49; H, 8.43; N, 6.54.

10 PREPARATION 2.l-(P-TOLI FENESULFONYI.)- HEXAMETHYLEN-EIMINE A mixtureof 9.92 g. of hexamethyleneimine, ml. of 2 N sodium hydroxide solution,and 18 g. of p-toluenesulfonyl chloride was shaken vigorously forfifteen minutes (heat of reaction noted) and allowed to stand for twohours. The product was recovered by filtration, washed with water, driedand then recrystallized from aqueous acetone anl gave 23.0 g. ofl-(p-toluenesulfonyl) hexamethyleneimine, M.P. 7l-73 C.

AnaIysia- CaIcd. for C H NO S (percent): C, 61.62; H, 7.56; S, 12.66.Found (percent): C, 61.83; H, 7.72; S, 12.94.

PREPARATION 3.-1-BENZOYLHEPTA METHYLENEIMINE A mixture of 30 g. ofheptamethyleneimine, 100 ml. of 50% sodium hydroxide solution, and 540ml. of water was cooled to 15 C. Benzoyl chloride (43 g.) was addeddropwise, with vigorous stirring and with the temperature maintained at15-20" C. Stirring was continued for 1 hour after the end of theaddition and the oily product recovered by ether extraction. Evaporationof the ether afforded 55 g. of 1-benzoylheptamethyleneimine as an oil.

PREPARATION 4.1-BENZOYLOCTA- M-ETHY-LENEIMINE A mixture of 10 g. ofoctamethyleneimine, 20 ml. of 50% sodium hydroxide and 100 ml. of water,was cooled to about 15 C. and 12.5 g. of benzoyl chloride was addeddropwise with vigorous stirring keeping the temperature 15-20 C.Stirring was continued for about 1 hour after the end of the benzoylchloride addition, and the product was recovered by methylene chlorideextraction. Evaporation of the solvent gave 17.6 g. ofl-benzoyloctamethyleneimine as an oil.

PREPARATION 5.1-BENZOYLALKYL- PIPERIDINES PREPARATION6.-1-B'ENZOYLDODECA- METHYLENEIMINE Azacyclotridecane-Z-one (40 g. in aSoxhlet thimble) was continuously extracted into a refluxing solution of15 g. of lithium aluminum hydride in one liter of ether. The reactionwas allowed to continue four hours after all of the amide had beenadded. Water was added to destroy the excess hydride and the ether phasewas separated, filtered through sodium sulfate, and concentrated invacuo to give 36.8 g. of oily dodecamethyleneimine. This was added to 30ml. of 50% sodium hydroxide in ml. of water, cooled to 20 C. with an icebath, and 30 g. of benzoyl chloride was added slowly with vigorousstirring. The resultant l-benzoy1dodecamethyleneimine precipitated fromsolution, was collected on a filter and Washed with water.Recrystallization from acetone-Skellysolve B gave 47.5 g. of1-benzoyldodecamethyleneimine, MP. 51- 53 C.; for analysis a sample wastwice recrystallized from acetone-Skellysolve B to M.P. 54-56 C.

Analysis.-Calcd. for C H NO (percent): C, 79.39; H, 10.17; N, 4.87.Found (percent): C, 79.43; H, 10.40; N, 5.03.

1 1 PREPARATION 7.BENZYL-3-AZABICYCLO- [3 .2.2] NONANE-3 -CARBOXYLATE Asuspension of 12.5 g. of 3-azabicyclo[3.2.2]nonane in 115 ml. of 2 Nsoduim hydroxide was stirred and chilled while adding 17 g. ofcarbobenzoxy chloride maintaining the temperature between 17-20 C. Themixture was stirred with no temperature control for one hour and thenextracted with ether. The extract was washed with dilute hydrochloricacid, water, dilute sodium bicarbonate, water, dried over sodiumsulfate, and the solvent removed to give a light yellow oil. The oilthus obtained was chromatographed over 500 g. of Plorisil (syntheticmagnesium silicate, hereinafter called Florisil) and eluted withSkellysolve B and Skellysolve B containing increasing proportions ofacetone (2 to 6% acetone). Those fractions which by thin layerchromatography were found to contain the desired product were combinedand evaporated to give 15.80 g. of benzyl-3-azabicyclo[3.2.2]nonane-3-carboxylate as a colorless oil.

Analysis.Calcd. for C H NO (percent): C, 74.10; H, 8.16; N, 5.40. Found(percent): C, 74.68; H, 8.22; N, 5.49.

PREPARATION 8.-3 P-TOLUENESULFO NYL) 3 -AZABICYC'LO[ 3 .2. 2] N ONANE Asuspension of 12.5 g. of 3-azabicyclo[3.2.2]nonane in 100 ml. of 2 Nsodium hydroxide was treated with 19.1 g. or p-toluenesulfonyl chloride.The mixture was shaken vigorously. No heat of reaction was noted so themixture was heated at the boiling point for several minutes, and cooledto room temperature. The lumpy product thus obtained was recovered byfiltration, washed with water, and recrystallized from acetone-water;yield 21.0 g. of 3-(ptoluenesulfonyl) 3 azabicyclo[3.2.2]nonane, M.P.115- 118 C.; an analytical sample from methanol melted at 115-117 C.

Analysis.-Calcd. for C H NO S (percent): C, 64.48;

H, 7.58; N, 5.01; S, 11.48. Found (percent): C, 64.20;

H, 7.76; N, 5.04; S, 11.53.

PREPARATION 9.-3 -BENZOYL-3 -AZABICYCLO [3.2.2] NONANE To a vigorouslystirred mixture of 500 g. of 3-azabicyclo[3.2.2]nonane and 3200 ml. of10% sodium hydroxide solution was added 500 ml. of benzoyl chloride.After minutes an additional 250 ml. of benzoyl chloride was added andstirring was continued for three hours. The product was recovered,washed with water and dried; yield 900 g. of 3-benzoyl 3azabicyclo[3.2.2]nonane, M.P. 92-95 C.; an analytical sample fromether-hexane melted at 93-94 C.

Analysis.--Calcd. for C H NO (percent): C, 78.56; H, 8.35; N, 6.11.Found (percent): C, 78.79; H, 8.52; N, 6.26.

PREPARATION l0.-2-BENZOYL-2AZABICYCLO [2.2.2] OCTANE To a stirredmixture of 50 g. of crude 2-azabicyclo [2.2.2]octane, 100 ml. of 50%sodium hydroxide solution, and 800 ml. of ice plus water was added 50ml. of benzoyl chloride. Stirring was continued for 1 hour after theaddition, and the crude product recovered by filtration and washed withwater. The solid product thus obtained was taken up in methylenechloride and filtered through about 100 n11. of Florisil, washed with 1l. of methylene chloride and then 1 l. of 10% acetone-Skellysolve B.Evaporation and recrystallization from acetone-Skellysolve B gave2-benzoyl-2-azabicyclo[2.2.2] octane, 33.8 g., M.P. 100-104 C. Foranalysis a sample was thrice recrystallized from the same solvent pairto M.P. 115- 118 C.

Analysis.-Calcd. for C H NO (percent): C, 78.10; H, 7.96; N, 6.51. Found(percent): C, 77.85; H, 7.22; N, 6.22.

Example 1.-0xygenation of l-benzoylpiperidine A medium was prepared of200 g. of cornsteep liquor (60% solids), g. of commercial dextrose, and10 l. of tap water. The pH was adjusted to between 4.8 and 5 and 10 ml.of lard oil was added as a foam preventive. This medium was sterilizedand inoculated with a 72-hour vegetative growth of Sporotrichumsulfurescens, ATCC 7159, and after incubation for 24 hours at atemperature of about 28 C. using a rate of aeration of 0.5 l. per minuteand agitation at 300 r.p.rn., the substrate, 2 g. of l-benzoylpiperidinein solution in a minimum amount of acetone (about 20 ml.) was then addedto the fermentation. After an additional 72-hour period of incubation atthe same temperature and aeration, the beer and mycelium were separatedby filtration. The mycelium was washed with water and the wash-water wasadded to the beer filtrate. The thus obtained beer filtrate wasextracted four times with a volume of methylene chloride equal toonefourth the volume of the filtrate. The combined extracts were washedwith one-fourth volume of distilled water and the solvent was removed bydistillation to give a residue comprising 1-benzoyl-4-hydroxypiperidine.

Chromatography of the extract residue over Florisil and elution withSkellysolve B containing increasing proportions of acetone gave 407 mg.of crude 1-benzoyl-4- hydroxypiperidine in the late 25%acetone-Skellysolve B eluate fractions. The product was characterized bythe following derivatives.

Oxidation of 151 mg. of the crude 4-hydroxy compound with Jones chromicacid reagent gave 137 mg. of crude 1-benzoyl-4-oxopiperidine whichafforded 89 mg. of the 2,4-dinitrophenylhydrazone, M.P. 196-198 C. Theremaining crude 4-hydroxy compound was treated with phenyl isocyanate togive, after chromatography on Florisil, 89 mg. of the phenylurethane,M.P. 184.5-186.5 C.

Example 2.Oxygenatio n of l-benzoylhexamethyleneimine The bioconversionand extraction procedures of Example l were repeated using 2 g. ofl-benzoylhexamethyleneimine as the substrate. The residue thus obtainedwas chromatographed on Florisil and eluted with Skellysolve B containingincreasing proportions of acetone. The 25% acetone-Skellysolve B eluategave about 250 mg. of 1- benzoyl-4-oxohexamethyleneimine and the acetoneeluate gave l-benzoyl 4 hydroxyhexamethyleneimine, determined by thinlayer chromatography.

The 1-benzoyl-4-hydroxyhexamethyleneimine thus obtained was dissolved inacetone and oxidized at room temperature by the addition of a visibleexcess of Jones reagent (2.67 M chromic acid reagent prepared from 26.7g. of chromium trioxide and 23 ml. of sulfuric acid diluted to 100 ml.with water). The excess oxidant was destroyed by the addition ofisopropyl alcohol and the mixture was evaporated to dryness. Water (20ml.) was added, and the product was extracted with 20 ml. of methylenechloride. The extract was evaporated to dryness and the residual1-benzoyl-4-oxohexamethyleneimine thus obtained was combined with thesame product obtained directly from the bioconversion. The combinedproduct was chromatographed on a column of Florisil. The column waseluted with Skellysolve B containing increasing proportions of acetoneand those fractions containing the desired product, determined by thinlayer chromatography were combined and evaporated to give about 770 mg.of l-benzoyl-4-oxohexamethyleneimine as an oil, B.-P. 170-174/0.3 torr,that crystallized slowly.

Analysis.Calcd. for C H NO (percent): C, 71.86; H, 6.96; N, 6.45. Found(percent): C, 71.51; H, 7.25; N, 6.46.

The 2,4 dinitrophenylhydrazone derivative ofl-benzoyl-4-oxohexamethyleneimine melted at 173.5180 C.

Example 3.-Oxygenation of l-benzoylhexamethyleneimine The bioconversion,oxidation, extraction and chromatographic procedures of Example 1 wererepeated on a larger scale using 5000 l. of sterilized medium of thesame composition and 1 kg. of l-benzoylhexamethyleneimine as thesubstrate to give 96.9 g. of 1-benzoy1-4-oxohexamethyleneirnine and12.57 g. of 1 benzoyl-3-oxohexamethyleneimine. The 3-oxo product wasrecrystallized three times from ethyl acetate-Skellysolve B hexanes togive 1-benzoyl-3-oxohexamethyleneimine, M.P. 113- 114 C.; 11 1705, 1625,11 1600, 1575, 1495; v, 785, 750, 705 cm.- in Nujol.

Analysis.Calcd. for C H NO (percent): C, 71.86; H, 6.96; N, 6.45. Found(percent): C, 71.48; H, 6.75; N, 6.49.

Example 4.Oxygenation of 1-(p-to1uenesulfonyl)- hexamethyleneimine Thebioconversion and extraction procedures of Example 1 were repeated using2 g. of l-(p-toluenesulfonyl)hexamethyleneimine as the substrate. Theresidue from the beer extract thus obtained was chromatographed over 100g. of Florisil by the gradient elution method. The residue was placed onthe Florisil with 75 ml. of methylene chloride; elution was with 4 l. ofSkellysolve B containing increasing proportions of acetone from 30% andcollecting fractions of 105 ml. each. The fractions which contained thedesired product, l-(p-toluenesulfonyl) 4 hydroxyhexamethyleneimine(determined by thin layer chromatography) were combined, evaporated toremove the solvent mixture, redissolved in acetone and oxidized withJones reagent in the same manner as disclosed in Example 2, above, togive a crude yellow solid, which was crystallized from ether to give0.33 g. l-(ptoluenesulfonyl)-4-oxohexamethyleneimine, M.P. 81 C.

Analysis.-Calcd. for C H NO S (percent): C, 58.40; H, 6.64; N, 5.24; S,12.00. Found (percent): C, 58.40; H, 6.47; N, 5.26; S, 12.24.

Recrystallization of the l-(p-toluenesulfonyl)-4-oxohexamethyleneiminethus obtained from acetone-Skellysolve B gave a crystallinemodification, M.P. 89-90 C.

Example 5.-Oxygenation of l-benzoylheptamethyleneimine The bioconversionand extraction procedures of Example 1 were repeated using1-benz0ylheptamethyleneimine as the substrate. The extract residue thusobtained, comprising a mixture of 1-benzoy1-4-hydroxyheptamethylimineand l-benzoyl-S-hydroxyheptamethyleneimine, was chromatographed onFlorisil. The hydroxylated l-benzoylheptamethyleneimines were elutedwith acetone and oxidized with excess Jones chromic acid reagent to thecorresponding ketoamides, which were separated by chromatography onFlorisil and eluted with Skellysolve B containing increasing proportionsof acetone. The more polar ketoamide was eluted with 25%acetone-Skellysolve B and recrystallized from acetone-Skellysolve B togive 1- benzoyl--oxoheptamethyleneirnine, M.P. 122-124" C.

Analysis.-Calcd. for C H NO (percent): C, 72.70; H, 7.41; N, 6.06. Found(percent): C, 72.63; H, 7.59; N, 6.31.

14 The less polar 1-benzoyl-4-oxoheptamethyleneimine was obtained as anoil from early 25% acetone-Skellysolve B eluate fractions.

Example 6.-Oxygenation of l-benzoyldodecamethyleneimine Thebioconversion and extraction procedures of Example 1 were repeated usingl-benzoyldodecamethyleneimine as the substrate. The extract residue thusobtained was analyzed by thin layer chromatography and found to containa mixture of hydroxy-l-benzoyldodecamethyleneimines, the majorcomponents of which were l-benzoyl-5 hydroxydodecamethyleneimine,1-benzoy1-6-hydroxydodecamethyleneimine and1-benzoy1-7-hydroxydodecamethyleneimine. The extract residue was thenchromatographed on Florisil and the mixedhydroxy-4-benzoyldodecamethyleneimines were eluted with acetone andoxidized in acetone with excess Jones reagent in the same manner asdisclosed in Example 2, above. The resulting mixture of ketoamides waschromatographed on Florisil. Elution with 25% acetone-Skellysolve B andthen, with acetone afforded, successively,l-benzoyl-S-oxododecamethyleneimine, 1-benzoyl-6oxododecamethyleneimine, and 1-benzoy1-7-oxododecamethyleneimine.

Example 7.-Oxygenation of 2-benzoyl-2-azabicyclo [2.2.2] octane Thebioconversion and extraction procedures of Example l were repeated using2 g. of 2-benzoyl-2-azabicyclo- [2.2.2]octane as the substrate.

The etxract residue thus obtained was chromatographed on Florisil.Elution with 25 acetone-Skellysolve B gave2-benzoyl-2-azabicyclo[2.2.2]octan-S-ol, which was recrystallized fromacetone-Skellysolve B to M.P. 139 141 C.

Analysis.--Calcd. for C H NO (percent): C, 72.70; H, 7.41; N, 6.06.Found (percent): C, 72.52; H, 7.19; N, 6.18.

The Z-benzoyl-S-hydroxy-Z azabicyclo[2.2.2]octane thus obtained isoxidized with Jones reagent in the manner disclosed in Example 2, aboveto obtain 2-benzoyl-2-azabicyclo [2.2.2] octan-5-one.

Example 8.Oxygenation of3-(p-toleuensulsulfonyl)-3-azabicyclo[3.2.2]nonane The bioconversion andextraction procedures of Example l are repeated using 2 g. of3-(p-toluenesulfonyl)- 3-azabicyc1o[3.2.2]nonane as the substrate.

The extract residue thus obtained is chromatographed on a column ofFlorisil. The column is eluted with Skellysolve B containing increasingproportions of acetone and those fractions which are shown to containthe desired products by thin layer chromatography are combined andevaporated to dryness to give3-(p-toluenesulfonyl)-3-azabicyclo[3.2.2]nonan-6-one and3-(p-toluenesulfonyl)-3-azabicyclo[3.2.2]nonan-6-ol.

The 3-(p-toluenesulfonyl)-3-azabicyclo[3.2.2]nonan-6- 01 thus obtainedis oxidized with Jones reagent in accordance with the proceduredisclosed in Example 2, above, to obtain3-(p-toluenesulfonyl)-3-axabicyclo [3.2.2]nonan-6-one.

Example 9.-Oxygenation of bispentamethyleneurea The bioconversion andextraction procedures of Example 1 were repeated using 5 g. ofbispentamethyleneurea as the substrate. The extract residue thusobtained was chromatographed on Florisil. Elution with Skellysolve Bcontaining increasing proportions of acetone and acetone gave1-pentamethylenecarbamyl-4-hydroxypiperidine in the 25acetone-Skellysolve B eluates and early acetone eluates. These fractionswere combined, evaporated to remove the solvent, redissolved in acetoneand oxidizd with excess Jones chromic acid reagent. The keto materialwas chromatographed on Florisil. Elution with 10% acetone-Skellysolve Band later 25 acetone- Skellysolve B afforded the desiredl-pentamethylenecarbamyl-4-oxopiperidine, which was recrystallized fromSkellysolve B several times to M.P. 66-67 C.

Analysis.--Calcd. for C N N O (percent): C, 62.83; H, 8.63; N, 13.32.Found (percent): C, 62.84; H, 8.36; N, 13.35.

Example 10.Oxygenation of l-benzoyl-Z-methyl- S-ethylpiperidine Thebioconversion and extraction procedures of Example 1 were repeated using2 g. of l-benzoyl-2-methyl-5- ethylpiperidine as the substrate. Theresidue thus obtained was chromatographed on a column of Florisil andthe column was eluted with Skellysolve B containing increasingproportions of acetone. The eluate fractions were analyzed by thin layerchromatography and those containing the desired product were combinedand the solvent removed by evaporation to give 1-benzoyl-2-methyl-4-hydroxy-5-ethylpiperidine.

The product thus obtained was oxidized with Jones reagent in the mannerdisclosed in Example 2, above, to give1-benzoyl-2-methyl-4-oxo-5-ethylpiperidine.

Example 11.xygenation of 3-benzoyl-3- azabicyclo[3.2.2]nonane A mediumwas prepared of 2.5 kg. of cornsteep liquor (60% solids), 1 kg. ofcommercial dextrose, and 125 l. of tap water, and adjusted to a pHbetween 4.8 and 5. This medium was sterilized, inoculated with a 72-hourvegetative growth of Sporotrichum sulfurescens, ATCC 7159, and incubatedfor 19 hours at a temperature of about 28 C. using a rate of aeration of6.25 l. per minute at about 300 r.p.m. The substrate, 25 g. of 3-benzoyl-3-azabicyclo[3.2.2]nonane in solution in a minimum amount ofacetone (about 250 ml.), was then added to the fermentation andincubation was continued for an additional 72-hour period under the sameconditions. Phenolized lard oil (5 g. phenol per liter of lard oil) wasadded as an antifoam agent as needed, about 50 ml. was used during thebioconversion. The beer and mycelium were then separated by filtrationand the mycelium was washed with water and the wash-water was added tothe beer filtrate. The thus obtained beer filtrate was extracted fourtimes with a volume of methylene chloride equal to one-fourth the volumeof the filtrate. The combined extracts were washed with onefourth volumeof deionized water and the solvent was removed by distillation atreduced pressure to give 25.8 g. of residue, which was analyzed by thinlayer chromatography and gas-liquid (vapor phase) chromatography andfound to contain unconverted substrate, 3-benzoyl-3-azabicyclo[3.2.2]nonane-6-one and 3-benzoyl-3-azabicyclo[3.2.2]nonane-6-ol. Gas chromatography showed that the residue containedunchanged substrate, 6-ketone, and 6- hydroxyl in the proportions of1:4.33:8.33.

The extract residue from the bioconversion (25.8 g.) was dissolved in500 ml. of acetone and oxidized by the addition of 32 ml. of Jonesreagent over a 5 minute period keeping the temperature of the solutionbelow 30 C. After the addition, the mixture was allowed to stir for tenminutes longer with no temperature control and excess oxidant was thendestroyed by the addition of 10 ml. of isopropanol. After adding 1 l. ofwater and stirring to dissolve sludge, the mixture was extracted threetimes with 250 ml. of methylene chloride each time. The combined extractwas washed once with 250 ml. of water, dried over sodium sulfate, andthe solvent removed by distillation, applying reduced pressure at thelater stage. The yield of crude residue as an oil was 23.2 g. Gaschromatography and thin layer chromatography showed the oil to be mainly3-benzoyl-3-azabcyclo[3.2.2]nonan- 6-one containing a small amount ofunconverted substrate. This material was dissolved in 250 ml. ofmethylene chloride and percolated through a bed of 175 g. of Florisil,eluting with 500 ml. of methylene chloride followed by 3000 ml. ofSkellysolve B containing 18% acetone. The combined eluate was taken todryness and the residual oil was distilled through a 4 inch Vigreuxcolumn at reduced pressure to give 12.17 g. of 3-benzoyl-3-azabicyclo[3.2.2]nonan-6-one B.P. 190-195/0.3 torr.

The following derivatives of 3-benzoyl-3-azabicyclo- [3.2.2]nonane-6-onewere prepared to confirm the structure:

Semicarbazone: M.P. 197-200 C.

Analysis.Calcd. for C H N O (percent): C, 63.98; H, 6.71; N, 19.65.Found (percent): C, 64.06; H, 6.82; N, 18.29.

Oxime: M.P. 156-158 C.

Analysis.Ca1cd. for C H N O (percent): C, 69.74; H, 7.02; N, 10.85.Found (percent): C, 69.71; H, 7.16; N, 10.87.

2,4Dinitrophenylhydrazone: M.P. l98201 C.

Analysis.-Calcd. for C H N O (percent): C,

59.56; H, 5.00; N, 16.54. Found (percent): C,

Example 12.0xygenation of benzyl-S-azabicyclo-[3.2.2]nonane-S-carboxylate The bioconversion and extraction proceduresof Example 11 were repeated using 50 g. of benZyl-3-azabicyclo-[3.2.2]nonane-3-carboxylate as the substrate. The beer extract residuethus obtained, containing benzyl-6-hydroxy-3-azabicyclo [3 .2.2]nonane-3-carboxylate, weighed 22.0 g. This was dissolved in 500 ml. ofacetone and 50 ml. of Jones reagent was slowly added while stiring andmaintaining the temperature between 25-30 C. during five minutes. Themixture was stirred ten mixtures longer at about 28 C. and excessoxidant was destroyed by the addition of 15 ml. of isopropanol. Most ofthe acetone was removed under reduced pressure, the mixture was dilutedwith 500 m1. of Water and extracted once with 200 ml. and four timeswith 100 ml. volumes of methylene chloride. The combined extract waswashed once with 250 ml. of water, dried over sodium sulfate, and thesolvent removed to give 19.1 g. of oil, which was chromatographed over500 g. of Florisil, placing it on the column with ml. of methylenechloride and eluting by the gradient method with 8 l. of Skellysolve Bplus increasing proportions of acetone from 0 to 20% and collectingfractions of about 375 ml. each. The fractions were examined byinfrared. Fractions 7-11 contained unchanged substrate. Fractions 14-20contained a ketone conversion product identified as one component byvaporphase chromatography. Fractions 14-20 containing the desiredproduct were combined and the solvent removed by distillation at reducedpressure to give 8.93 g. ofbenzyl-6-oxo-3-azabicyclo[3.2.2]nonane-3-carboxylate as a yellow oil.

Analysis.-Calcd. for C I-I NO (percent): C, 70.31; H, gel; N, 5.13.Found (percent): C, 70.58; H, 7.41; N,

The 2,4-dinitrophenylhydrazone derivative of the henzyl 6 oxo 3azabicyclo[3.2.2]nonane 3 carboxylate melted at 84-86 C.

Analysis.Calcd. for C H N O (percent): C, 58.27; H, 5.11; N, 15.45.Found (percent): C, 58.43; H, 5.21; N, 15.32.

Example 13.-Oxygenation of 1-benzoyloctarnethyleneimine Thebioconversion and extraction procedures of Example 11 were repeatedusing 15 g. of l-benzoyloctamethyleneimine as the substrate. The extractresidues thus obtained, containing mainly1-benzoyl-4-hydroxyoctamethyleneimine and1-benzoyl-S-hydroxyoctamethyleneimine, was taken up in acetone (ca. 1liter) and oxidized with excess Jones reagent. The resultant ketoamidemixture was chromatographed on Florisil. The less polar ketoamide,4-oxo-1-benzoyloctamethyleneimine, was eluted with 25%acetone-Skellysolve B, and the more polar ketoamide,

1 7 5-oxo-1-benzoyloctamethyleneimine, was eluted with acetone.

Example 14.--xygenation of l-benzoyloctamethyleneimine The bioconversionand extraction procedures of Example 11 were repeated using 25 g. ofl-benzoyloctamethyleneimine as the substrate. The extract residue thusobtained was dissolved in 500 ml. of acetone. The solution was cooled onan ice bath. Excess Jones reagent was added dropwise to the solution.After 10 minutes, the excess Jones reagent was decomposed by theaddition of isopropyl alcohol. The mixture was concentrated to an oilyresidue, which was dissolved in 200 ml. of water and 100 ml. ofmethylene chloride. The aqueous phase was separated and extracted withmore methylene chloride (2x 100 ml.). The combined extracts were driedand concentrated. The residual material thus obtained was dissolved in100 ml. of methylene chloride and the solution was placed on a 2.5 kg.column of Florisil packed in Skellysolve B hexanes. The following twoliter fractions were collected: 2 of Skellysolve B, 10 of 10% (v./v.)acetone in Skellysolve B, 8 of 20% acetone in Skellysolve B, 5 or 50%acetone-Skellysolve B, and 3 of acetone. Fractions 15 through 18 gave4.80 g. of 1-benzoyl-4-oxooctamethyleneimine as a crystalline solid andfractions 20 through 23 gave 11.62 g. ofl-benzoyl-S-oxooctamethyleneimine as a crystalline solid. A sample (1.60g.) of the 1-benzoyl-4oxooctamethyleneimine was dissolved in methylenechloride and placed on a column of Florisil (8 0 g.) packed withSkellysolve B. Elution with 10% (v./v.) acetone in Skellysolve B gavecolorless crystals. Two recrystallizations from cold methylene chloride-Skellysolve B gave 1-benzoyl-4-oxooctamethyleneimine as colorlessplates, M.P. 87-88 C.; 1 1700 m.s., 1625 s., v, 787 m.s., 744 m.s., 705m.s.; cm.- in Nujol.

Analysis.-Calcd. for C H NO (245.31) (percent): C, 73.44; H, 7.81; N,5.71. Found (percent): C, 73.30; H, 8.03; N, 5.80.

A sample (11.40 g.) of the l-benzyl-5-oxooctamethyleneimine dissolved inmethylene chloride was placed on a column of Florisil (600 g.) packedwith Skellysolve B. Elution with 25% (v./v.) acetone in Skellysolve Bgave colorless crystals, which were recrystallized from acetone-Skellysolve B to M.P. 69-71 C. A final recrystallization fromacetone-Skellysolve B gave l-benzoyl-S-oxooctamethyleneimine ascolorless crystals, M.P. 7072 C.; 11 1700 m.s., 1625 s., v, 800 m., 748m.s., 717 s. cm in Nujol.

Analysis.-Calcd. for C H NO (percent): C, 73.44;

H, 7.81; N, 5.71. Found (percent): C, 73.54; H, 7.72;

Example 15.Oxygenation of 2-benzoyl-2-azabicyclo [2.2.2]octane Thebioconversion and extraction procedures of Example 11 were repeatedusing 25 g. of 2-benzloyl-2-azabicyclo[2.2.2]octane as the substrate.The extract residue thus obtained was dissolved in methylene chlorideand chromatographed over 1000 g. of Florisil. Elution was with 4 1. eachof Skellysolve B hexanes containing 10%, 15%, acetone and 12 l. ofSkellysolve B containing acetone; cuts of 800 ml. each were collected;pooled residues as follows:

(A) Fractions 7-11=3.98 g. unchanged substrate (B) Fractions 17l8=l.61g. of 2-benzoyl-2-azabicyclo [2.2.2]octan-exo-6-ol (C) Fraction 19=2.14g. of mixture 18 (D) Fractions 20-27=12.16 g. of2-benzoyl-2-azabicyclo[2.2.2]octan-exo-S-ol B above was recrystallizedfrom acetone; yield, 1.61 g. of2-benzoyl-2-azabicyclo[2.2.2]octan-exo-6-ol, M.P. 200- 205 C.

Analysis.-Calcd. for C H NO percent): C, 72.70; H, 7.41; N, 6.03. Found(percent): C, 72.70; H, 7.64; N, 5.82.

D above was recrystallized from acetone; yield, 10.62 g. of2-benzoyl-2-azabicyclo[2.2.2]octan-exo-5-ol, M.P. 146148 C.

Analysis.-Calcd. for C H NO (percent): C, 72.07; H, 7.41; N, 6.03. Found(percent): C, 72.52; H, 7.19; N, 6.18.

Example 16.Oxygenation of 3-benzoyl-3-azabicyclo [3.3.1]nonane Thebioconversion and extraction procedures of Example 11 were repeatedusing 25 g. of 3-benzoyl-3-azabicyclo[3.3.1]nonane as the substrate. Theextract residue thus obtained was chromatographed on Florisil (2.0 kg.)packed with Skellysolve B. Elution with 25% and 50% (v./v.)acetone-Skellysolve B gave crystalline material (19.645 g.).Recrystallization from acetone preceded by decolorization with activatedcharcoal gave colorless crystals, M.P. 139-141 C. From the several cropscollected, a total of 16.894 g. (0.0689 mole, 63%) of 3-benzoyl-3-azabicyclo[3.3.1]nonan-axial-6-ol was obtained, M.P. 137-141 C. Threerecrystallizations from acetone gave 3 benzoyl 3azabicyclo[3.3.1]nonan-axial 6 o1, M.P. 139-141 C.; [a1 1 (c., 0.860,chloroform); v 3460, 3420 sh., 1 c=c 1610, 1575,1525, 1495, v, 785, 740,705 cm." in Nujol.

Analysis.Calcd. for C H NO (percent): C, 73.44; H, 7.81; N, 5.71. Found(percent): C, 73.66; H, 7.96; N, 607.

3 benzoyl 3 azabicyclo[3.3.l]nonane was bioconverted in accordance withthe procedure described above, using the microorganism Rhizopusarrhizis, ATTC 11145 (American type culture collection), to give3-benzoyl-3- azabicyclo[3.3.1]nonan-1-ol,

M.P. 138-140 C.; [ab-40 (c., 0.803, chloroform); v 3330, 1 6:0 1600,1575, 1520, 1495, v, 725, 705 cm" in Nujol.

Analysis.--Calcd. for C H N0 (percent): C, 73.44; H, 7.81; N, 5.71.Found (percent): 73.46; H, 7.76; N, 6.15.

and 3 benzoyl axial 6 hydroxy 3 azabicyclo[3.3.1] nonane, M.P. -137 0.,having an infrared spectrum identical with that of the hydroxylatedproduct isolated from bioconversion with Sporotrichum sulfurescens,above.

Example 17.-0xygenation of 1-benzoy-2- propylpiperidine Thebioconversion and extraction procedures of Example 11 were repeatedusing 25 g. of l-benzoyl-Z-propylpiperidine as the substrate. Theextract residue thus obtained was dissolved in methylene chloride andchromatographed on Florisil to give 0.524 g. of 1benzoyl-4-hydroxy-2-propylpiperidine which was recrystallized fromacetone-hexanes to M.P. 124130 C. (0.382 g.)

Analysis.-Calcd. for C H NO (percent): C, 72.84; H, 8.56; N, 5.66. Found(percent): C, 72.64; H, 8.71; N, 5.44.

and 0.674 g. of 1-benzoyl-2-(2-hydroxypropyl) piperidine which wasrecrystallized from acetone-hexanes to M.P. 116-119" C.

Analysis.Calcd. for C H NO (percent): C, 72.84; H, 8.56; N, 5.66. Found(percent): C, 73.09; H, 8.47; N, 5.59.

A 0.157 g. sample of the 1-benzoyl-4hydroxy-2-propylpiperidine wasdissolved in acetone and oxidized with excess Jones reagent to give1-benzoyl-4-oxo-2-propylpiperidine.

A 0.160 g. sample of the 1-benzyl-2-(2-hydroxypropyl) piperidine wasdissolved in acetone and oxidized with excess Jones reagent to give1-benzoyl-2-(2-oxopropyl) piperidine.

Example 18.Oxygenation of 1-benzoyl-4-propylpiperidine The bioconversionand extraction procedures of Example 11 were repeated using 25 g. of1-benzoyl-4-propylpiperidine as the substrate. The extract residue thusobtained was chromatographed on Florisil and eluted with hexanescontaining increasing proportions of acetone. The appropriate fractionsdetermined by NMR and IR were combined and distilled to give1-benzoy-4-(2-oxopropyl) piperidine, B.P. 168-172 C. (0.15 mm.) N1.5441.

Example 19.Oxygenation of 1-benzoyl-2-methylpiperidine The bioconversionand extraction procedures of Example 11 were repeated using 25 g. ofl-benzoyl-2-methy1- piperidine as the substrate. The extract residuethus obtained was charomatographed on Florisil. The fractions containingthe 4-oxo product were combined and recrystallized from acetone-hexanesto give 0.252 g. which was recrystallized again from acetone-hexanes togive 1- benzoyl-4-oxo-2-methylpiperidine, M.P. 117-118 C.

Analysis.-Calcd. for C H NO (percent): C, 71.86; H, 6.96; N, 6.45. Found(percent): 7, 72.21; H, 7.07; N, 6.46.

The fractions from the chromatogram which contained the 3-hydroxyproduct were combined and recrystallized from acetone-hexanes to give1.225 g. of product which was twice recrystallized from acetone-hexanesto give 1- benzoyl-3-hydroxy-2-methylpiperidine, M.P. 127-129 C.; 11)

Analysis.-Calcd. for C H NO (percent): C, 71. H, 7.82; N, 6.39. Found(percent): C, 71.26; H, 8.00 N, 6.68.

The fractions containing the 4-hydroxy product were combined andrecrystallized from acetone-hexanes to give 4.277 g. of product whichwas recrystallized from acetone-hexanes to give1-benzoy1-4-hydroxy-2-methylpiperidine, M.P. 124-125 C.; [ab 29.

Analysis.-Calcd. for C H NO (percent): C, 71.20; H, 7.82; N, 6.39. Found(percent): C, 71.25; H, 7.83; N, 6.65.

A 0.352 g. sample of the l-benzoyl 3 hydroxy 2- methylpiperidine in 15ml. of acetone was oxidized with excess Jones reagent and concentratedon a steam bath. Water was added and the solution was extracted withchloroform (3x 15 ml.). The combined extracts were dried of magnesiumsulfate and concentrated under reduced pressure to an oil which wascrystallized from methylene chloride-hexanes to give 0.163 g. ofl-'benzoyl- 3-oxo 2 methylpiperidine, M.P. 102-104 C., which wasrecrystallized to give colorless plates, M.P. 103-105 C.; [ab +74.

Analysis.Calcd. for C H NO (percent): C, 71.86; H, 6.96; N, 6.45. Found(percent): C, 71.99; H, 7.09; N, 6.25.

A 0.5 g. sample of l-benzoyl 4 hydroxy-Z-methylpiperidine was dissolvedin 25 ml. of acetone, oxidized and worked up in the manner describedabove to give 0.325 g. of l-benzoyl 4 oxo 2 methylpiperidine, M.P. 116-119 C. The infrared spectrum was identical to the same material obtaineddirectly from the bioconversion, above.

20 Example 20.-Oxygenation of l-benzoyl-4-methylpiperidine Thebioconversion and extraction procedures of Example 11 were repeatedusing 25 g. of 1-benzoyl-4-methylpiperidine as the substrate. Theextract residue thus obtained was chromatographed on Florisil. Thefractions containing the 4-hydroxy product were combined andrecrystallized from acetone-hexanes to give 1.342 g. which wasrecrystallized twice again from acetone-hexanes to give1-benzoyl-4-hydroxy-4-methylpiperidine, M.P. 104- 106 C.

Analysis.Calcd. for C H NO (percent): C, 71.20; H, 7.82; N, 6.39. Found(percent): C, 71.24; H, 7.77; N, 6.53.

The fractions from the chromatogram which contained the hydroxy methylproduct were combined and recrystallized from acetone-hexanes to give4.721 g. of product which was twice recrystallized from acetone-hexanesto give l-benzoyl 4 hydroxymethylpiperidine, M.P. 92- C.

Analysis.-Calcd. for C13H17NO2 (percent): C, 71.20; H, 7.82; N, 6.39.Found (percent): C, 71.37; H, 7.94; N, 6.43.

Example 21 .3-benzoyl-3-azabicyclo [3 .3 .1] nonan-6-one A solution of3-benzoyl 3 azabicyclo[3.3.1]nonanaxial-6-ol (1.543 g., 0.00630 mole) inacetone (150 ml.) was cooled on an ice bath and treated with an excess(1.8 ml.) of Jones reagent. After 30 minutes at room temperature, theexcess oxidant was consumed with isopropyl alcohol. The solution wasdecanted and filtered through sodium sulfate. The residue was dissolvedin water and extracted with methylene chloride. The combined organicsolutions were dried over magnesium sulfate, 'Celite was added, and themixture again filtered. The filtrate was concentrated under reducedpressure and cooled. Crystallization gave 0.882 g. (0.00363 mole, 57%)of product, M.P. 158-160 C. Two recrystallizations from acetone gavecolorless crystals of 3-benzoyl- 3 azabicyclo[3.3.1]nonan 6 one, M.P.159-161 C.; 11 1705, 1620, 11 1600, 1580, 1570, 1490, v, 785, 775, 735,700 cm:- in Nuqol.

Analysis.-Calcd. for C H NO (percent): C, 74.05; H, 7.04; N, 5.76. Found(percent): C, 73.16; H, 7.30; N, 5.29.

Example 22.2-benzoyl-2-azabicyclo[2.2.2]octan-5-one Two grams of2-benzoyl-2-azabicyclo[2.2.2]octan-5-ol dissolved in ml. of acetone wastreated dropwise with a slight excess of a stock solution of oxidizingreagent prepared from 26.7 g. of chromium trioxide, 50 m1. of water, and23 ml. of concentrated sulfuric acid (Jones reagent). An ice bath wasapplied as necessary to keep the temperature below 30 during theoxidation. Isopropanol (5.0 ml.) was added to destroy excess chromicacid. The mixture was diluted with 200 ml. of water and extracted withmethylene chloride. The extract was washed once with water and driedover sodium sulfate. The solvent was removed by distillation andresulting product crystallized to yield 1.95 g. of2-benzoyl-2-azabicyclo[2.2.2]octan-5-one, M.P. 66-72 C.; IR (CHC1solution), 1740 cm.- (C=O).

Example 23.-2-benzoyl-2-azabicyclo [2.2.2] octan-6-one A mixture of 300mg. of 2-benzoyl-2-azabicyclo[2.2.2] octan-6-ol (300 mg.) in 20 ml. ofacetone was oxidized with chromic acid in the manner described inExample 22, above. The product thus obtained was recrystallized fromacetone-hexane to give Z-benzoyl 2 azabicyclo [2.2.2]octan-6-one M.P.99-101 C.; IR (CHCI soluhon), 1740 cmr (C=O).

21 Example 24.-1-benzoyl-5-hydroxyheptamethyleneimine A solution of 700mg. of 1-benzoyl-S-oxoheptamethyleneimine (from Example 5, above) in 12ml. of methanol was reduced with a solution of 150 mg. of sodiumborohydride in 2 ml. of N/ 10 sodium hydroxide at room temperatureovernight. The product was recovered by acidification with acetic acid,evaporation of most of the methanol at reduced pressure, and partitionbetween Water and methylene chloride. Chromatography of the organicsolution of Florisil gave 657 mg. of1-benzoyl-5-hydroxyheptamethyleneimine in the acetone eluate; foranalysis the material was recrystallized from acetone-Skellysolve B toM.P. 116-118 C.

Analysis.Calcd. for C H NO (percent): C, 72.07; H, 8.21; N, 6.00. Found(percent): C, 72.18; H, 8.01; N, 6.03.

In the same manner following the procedure of Example 23, other ketocompounds of this invention can likewise be reduced to the correspondinghydroxy compounds by substituting the appropriate keto compound asstarting material in place of l-benzoyl 5 oxoheptamethyleneimine. Thefollowing conversions are representative:

1-benzoy1-4-oxoheptamethyleneimine to 1-benzoyl-4-hydroxyheptamethyleneimine;1-(p-toluenesulfonyl)-4-oxohexamethyleneimine to 1-(p-toluenesulfonyl)-4-hydroxyhexamethyleneimine;l-benzoyl-S-oxododecamethyleneimine to l-benzoyl-S-hydroxydodecamethyleneimine; l-benzoyl-6-oxododecamethyleneimine tol-benzoyl-6- hydroxydodecamethyleneimine;1-benzoyl-7-oxododecamethyleneimine to l-benzoyl-7-hydroxydodecamethyleneirnine; l-benzoyl-4-oxooctamethyleneimine to1-benzoy1-4- hydroxyoctamethyleneimine;1-benzoyl-5-oxooctamethyleneimine to l-benzoyl-S-hydroxyoctamethyleneimine; 3- (p-toluenesulfonyl) -3-azabicyclo[ 3.2.2]nonan-6- one to 3-(p-toluenesulfonyl)-3-azabicyclo- [3.2.2]nonan-6-ol;

and the like.

Example 25.-3-benzoyl-3-azabicyclo[3.3.1] nonan-equatorial-6-ol Asolution of sodium boroh'ydride (1.0 g., 0.0265 mole) in 10 ml. of 0.1 Maqueous sodium hydroxide was added to a solution of 3 benzoyl 3azabicyclo[3.3.1] non-6-one (1.017 g., 4.18 mmoles) in 40 ml. ofmethanol. Thin layer chromatography after 0.5 hour showed the reactionto be complete. The solution was partially concentrated under reducedpressure and then was diluted with 150 ml. with water. The solution wasmade acidic (pH 5-6) with acetic acid and was concentrated under reducedpressure over a hot water bath until crystals began to form. The mixturewas extracted with methylene chloride (3 X 50 m1.). From the dried(magnesium sulfate) extract solution, an oil was obtained followingconcentration. 'Ihe oil crystallized and the solid was recrystallizedfrom acetone-Skellysolve B, giving 0.695 g. (2.84 mmoles, 67%) of 3benzoyl-3-azabicyclo[3.3.1]nonan-equatorial-6-ol, M.P. 135-l38 C., whichwas recrystallized twice from acetone-Skellysolve B to M.P. 139-141 C.;3360, 11 1600, who 1590, 1575, 1530, 1490, 11 1060, 11 790, 780, 735,705 cm." in Nujol.

Analysis.Calcd. for C H NO (percent): C, 73.44; H, 7.81; N, 5.71. Found(percent): 0, 73.92; H, 7.95; N, 6.08.

Example 26.3-benzoyl-3-azabicyclo[3.2.2] nonan-exo-6-ol Twenty grams ofthe crude 3 benzoyl 3 azabicyclo [3.2.2]nonan-exo- 6-one dissolved in350 ml. of methanol was treated with a solution of 16.0 g. of sodiumborohydride in ml. of N/ 10 sodium hydroxide for 30 minutes. Thin layerchromatography indicated complete reaction. The mixture was diluted with300 ml. of water and allowed to stand for 18 hours with chilling. Themixture was then adjusted to pH 6 by the cautious addition of 50% aceticacid. The solid product thus obtained was recovered by filtration,washed with water and dried; yield, 12.75 g. of 3 benzoyl3-azabicyclo[3.2.2]nonan-exo-6-ol, M.P. 131-135 C.; an analytical samplefrom acetone melted at 135-137 C.

Analysis.Calcd. for C15H19H19No2 (PI'Cent): C, H, 7.81; N, 5.71. Found(percent): C, 73.31; H, 7.87;

Example 27.3-azabicyclo[3.2.2]nonan-exo-6-ol 6-benz0ate hydrochloride(acyl migration) To a warm solution of 3-benzoyl-3-azabicyclo[3.2.2]nonan-exo-6-ol (2.14 g.) in 50 ml. of tetrahydrofuran was added 4.0 ml.of concentrated hydrochloric acid and the mixture was allowed to standat 25 C. The solution was examined at intervals by thin layerchromatography and after complete reaction (23 hours), it wasconcentrated under reduced pressure. The oil thus obtained wastriturated twice with ether, decanting 01f the ether each time, and theHCl salt was precipitated by adding 25 ml. of acetone and 50 ml. ofether; yield, 2.16 g., M.P. 205-208 C. This was recrystallized frommethanol-methyl ethyl ketone; yield 2.06 g. of3-azabicyclo[3.2.2]nonan-exo-6-ol 6-benzoate hydrochloride, M.P. 205-208C.; IR amine, HCl at 2250-2700 cm; ester C=O at 1710 cm.-

Analysis.Calcd. for C H NO Cl (percent): C, 63.93; H, 7.15; N, 4.97.Found (percent): C, 63.92; H, 7.44; N, 5.24.

Example 28.-3-benzoyl-3-azabicyclo [3.3.1 nonan-o-one ethylene ketal Amixture of 3-benzoyl-3-azabicyclo[3.3.1]nonan-6-one (0.550 g., 0.00226mole) in benzene (100 ml.), p-toluenesulfonic acid hydrate (0.090 g.,0.000473 mole), and ethylene glycol (10 ml.) was heated to reflux for 18hours. The condensate was dried by passing through a calcium carbidetrap. A few drops of pyridine were added and the mixture was cooled toroom temperature. The mixture was extracted with 5% aqueous sodiumbicarbonate solution (50 ml.) and with water (2X 25 ml.). The benzenelayer was dried over magnesium sulfate and concentrated under reducedpressure to give as 3-benzoyl-3-azabicyclo- [3.3.1]nonan-6-one ethyleneketal as an oil, v 1630, v 1605, 1580, 1500, 11 708 emf- Example29.-8-benzoyl-1,4-dioxa-8-azaspiro- [4.7] dodecane A mixture of 1benzoyl 4 oxoheptametheneimine (16.487 g., 0.0712 mole),p-toluenesulfonic acid hydrate (1.35 g., 0.00710 mole), ethylene glycol(25 ml.), and benzene (200 ml.) was heated to reflux for 24 hours. Thecondensate was passed through a calcium carbide drying trap during thistime. Pyridine (2.0 ml.) was added and the mixture was cooled. Themixture was extracted with aqueous sodium bicarbonate solution and withwater (3 X 100 ml). The benzene layer was dried over magnesium sulfateand concentrated to give 8-benzoyl-l,4-dioxa-8- azaspiro[4.7]dodecane asa viscous oil.

Example 3 0.--9-benzoyl-1,4-dioxa-9-azaspiro- [4.7] dodecane A mixtureof l-benzoyl-S-heptamethyleneimine (39.883 g., 0.0172 mole),ptoluenesulfonic acid (2.96 g., 0.0155 mole), ethylene glycol (75 ml.,83 g., 1.34 mole), and benzene (500 ml.) was heated to reflux. Thecondensate was dried by passing it through a calcium carbide trap. Themixture was refluxed 30 hours. Pyridine (6.0 ml.) was then added to thecooled mixture, the benzene layer was extracted with 5% aqueous sodiumbicarbonate (3X 100 ml.) and dried over magnesium sulfate. Concentrationof the benzene solution gave 42.95 g. (0.156 mole, 90% yield) of aviscous oil which was crystallized from ether to give9-benzoyl-1,4-dioxa-9-azaspiro [4.7]dodecane, M.P. 7274 C. Tworecrystallizations from ether-Skellysolve B gave colorless crystals of9-benzoyl-1,4-doxa-9-azaspiro- [4.7]dodecane, M.P. 72-73 C., 11 1630 s.cm.- in Nujol.

Analysis.Calcd. for C H NO (275.34) (percent): C, 69.79; H, 7.69; N,5.09. Found (percent): C, 69.99; H, 7.84; N, 5.20.

Example 3 1.-9-benzoyl-1,4-dioxa-9-azaspiro- [4.8] tridecane A solutionof 1 benzoyl-S-oxooctamethyleneimine (18.819 g., 0.0768 mole) andp-toluenesulfonic acid hydrate (1.46 g.) in benzene (200 ml.) withethylene glycol (30 ml.) was heated to the reflux temperature of benzenefor 22 hours and the condensate was passed through a calcium carbidedrying trap. Pyridine (3.0 ml.) was added at the end of the refluxperiod, the mixture was cooled and then extracted with 5% aqueous sodiumbicarbonate (100 ml.) and with water (2x 100 ml.). The benzene layer wasdried over magnesium sulfate, and concentrated under reduced pressure,giving a viscous yellow oil. The infrared spectrum of the oil showed thepresence of a carbonyl function (1730 w. cm.- and so the oil wasresubjected twice to the ketalization process. The crude oily productcrystallized spontaneously on standing. Recrystallization fromether-Skellysolve B gave two crops (13.682 g., 0.0473 g., 0.0473 mole,61%) of light yellow crystals, M.P. 90-95" C., which were recrystallizedfrom ether- Skellysolve B to give 9-benzoyl-1,4-dioxa-9-azaspiro[4.8]tridecane, M.P. 98-100 C. A final recrystallization preceded bydecolorization with activated charcoal gave colorless needles of9-benzoy1-1,4-dioxa-9 azaspiro[4.8] tridecane, M.P. 99-101" C., 11 1630s., v, 798 m., 737 m., 704 m.s. cm.- in Nujol.

Analysis.Calcd. for C17H23NO'3 (289.36) (percent): C, 70.56; H, 8.01; N,4.84. Found (percent): C, 70.50; H, 8.08; N, 5.16.

Example 32.-8-benzoyl-l,4-dioxa-8-azaspiro[4.6] undecane A mixture of11.957 g. (0.0551 mole) l-benzoyl-4-oxohexamethyleneimine in 200 ml. ofbenzene, 1.020 g. (0.00537 mole) of p-toluenesulfonic acid hydrate and28 ml. of ethylene glycol was stirred and heated at reflux for about 24hours. The condensate was passed through a calcium carbide trap toremove water. Pyridine (3 ml.) was added at the end of the reflux periodand the mixture cooled. The mixture was extracted with 5% aqueousbenzene layer was dried and concentrated under reduced pressure to give8-benzoyl-1,4-dioxa 8 azaspiro[4.6]- undecane as an oil, 11 1630 cm.-

Example 33.-8-benzyl-1,4-dioxa-8 azaspiro[4.6] undecane The product fromExample 32, above, was dissolved in 75 m1. of ether and added dropwiseto a mixture of lithium aluminum hydride (2.0 g.) and ether (200 ml.).The mixture was stirred and heated to reflux for 3 hours, stirred atroom temperature for about 72 hours, and again heated to reflux for 2hours before the excess lithium aluminum hydride was decomposed by theaddition of ethyl acetate and water. The solids thus obtained wereremoved by filtration and were washed with ether. The filtrate andwashes were combined, dried and concentrated to an oil which wasdistilled to give 9.696 g. (0.0392 mole, 71%) of 8-benzyl-1,4-dioxa-8-azaspiro [4.6]undecane as a pale yellow oil, B.P. 120-121 C.

(0.13 mm.); n 1.5312; v 1600, 1580, 1490, v, 730, 695 cmr'Analysis.-Calcd. for C H NO (247.33) (percent): C, 72.84; H, 8.56; N,5.66. Found (percent): C, 73.22; H, 8.98; N, 6.11.

Example 34.-9-benzyl-1,4-dioxa-9-azaspiro [4.7]dodecane A solution of 9benzoyl 1,4 dioxa 9 azaspiro [4.7]dodecane (40.9 g., 0.148 mole) inether (300 ml.) was added slowly to a stirred mixture of lithiumaluminum hydride (6.0 g., 0.158 mole) and ether (200 ml.). The resultingmixture was stirred at room temperature for 16 hours and at refluxtemperature for 5 hours. The excess lithium aluminum hydride wasdecomposed with 1:1 acetone-water and with water. The inorganic solidswere collected on a filter and washed three times with ether. Thecombined ether solution was dried and concentrated to an oil (38.26 g.).Simple distillation gave 9-benzyl- 1,4-dioxa-9-azaspiro[4.7]dodecane asa colorless oil (33.72 g., 0.129 mole, 87% yield), B.P. -127 C. (0.3mm.); v 1115 s. cmr- Analysis.Calcd. for C H NO (261.35 (percent): C,73.53; H, 8.87; N, 5.36. Found (percent): C, 73.39; H, 8.31; N, 5.34.

Example 35 .8-benzyl-1,4-dioxa-8-azaspiro [4.7]dodecane The8-benzoyl-1,4-dioxa-8-azaspiro[4.7]dodecane from Example 30, above, wasdissolved in ether (2 l.) and slowly added to a stirred mixture oflithium aluminum hydride (3.0 g., 0.0790 mole) and ether (200 ml.). Theexcess ether was allowed to boil off. The remaining oil was dissolved intetrahydrofuran and added to the re action mixture. The mixture wasstirred and refluxed 7 hours and then kept at room temperatureovernight. The excess lithium aluminum hydride was destroyed by additionof ethyl acetate and water. The inorganic solids were removed byfiltration and were Washed with ether. The combined ether filtrates werecombined, dried, and concentrated to a yellow oil. Distillation of theoil gave 8-benzyl- 1,4-dioxa-8-azaspiro[4.7]dodecane (5.700 g., 0.0218mole, 30% yield) as colorless oil, B.P. -150 C. (0.2 mm.); 11 1.5284; v1600 m.w., 1580 w., 1490 s. v, 7.25 s., 695 s. cm.

Analysis-Calcd. for C H NO (261.35) (percent): C, 73.53; H, 8.87; N,5.36. Found (percent): C, 73.33; H, 8.91; N, 5.14.

Example 36.-9-benzyl1-1,4-dioxa-9-azaspiro [4.8]tridecane A solution of9-benzoyl-1,4-dioxa-9-azaspiro [4.8]tridecane (13.331 g., 0.0462 mole)in ether (250 ml.) was dribbled into a mixture of lithium aluminumhydride (2.0 g.) in ether (100 ml.). The mixture was stirred at roomtemperature for 16 hours and at reflux temperature for 4 hours. Theexcess hydride was decomposed by the cautious addition of water. Theinorganic solids were removed by filtration and washed with ether. Thecombined ether solution was dried and concentrated to an oil. A simpledistillation of the oil gave 9-benzyl-1,4-dioxa-9-azaspiro[4.8]tridecane (10.779 g., 0.0392 mole, 85% yield) as a colorless oil,B.P. -147 C. (1.5 mm.); n 1.5302; v 1600 w., 1498 m., r, 750 m., 715m.s., 701 m.s. cmr

Analysis.-Calcd. for C H NO (275.38) (percent): C, 74.14; H, 9.15; N,5.09. Found (percent): C, 74.77; H, 9.66; N, 5.04.

The product crystallized when kept overnight in the refrigerator, M.P.44-46 C.

25 Example 37.-3-benzyl-3-azabicyclo[3.3.1]nonan- 6-one ethylene ketal Asolution of the 3-benzoyl-3-azabicyclo[3.3.1]nonan- 6-one ethylene ketalfrom Example 29, above, in ether was reduced with lithium aluminumhydride (0.5 g.) in ether. After refluxing 4 hours, the excess hydridewas decomposed with ethyl acetate and water, the solids were filteredoff, the ether solution was dried over magnesium sulfate andconcentrated to give 3-benzyl-3-azabicyclo [3.3.1]nonan-6-one ethyleneketal as an oil; VCH 2930, 2900, 2875, v 1600, 1580, 1495, v, 736, 700cmr Example 38.--3-benzyl-3-azabicyclo[3 .3. 1 nonan- 1-01 A suspensionof 3- benzoyl-1-hydroxy 3 azabicyclo [3.3.1]nonane (0.478 g., 0.00195mole) in ether was added to a mixture of lithium aluminum hydride (0.6g.) and ether. The resulting mixture was stirred at room temperatureovernight and then heated to reflux temperature for 7 hours. The excesshydride was decomposed by addition of ethyl acetate and Water. Thesolids were removed by filtration through celite and the ether solutionwas dried (magnesium sulfate). An oil was obtained upon re moval of theether. The oil crystallized in the cold and crystals formed from a coldpentane solution and were collected giving 0.206 g. (0.00892 mole, 45%)of crystals, M.P. 74-76 C. Recrystallization, preceded by decolorizationwith activated charcoal gave colorless crystals of 3-benzyl-3-azabicyclo[3.3.1]nonan 1 ol, M.P. 7376 C.; [ab 24 (C, 0.740,CHCl 1 3280, 3210, v

1600, 1585, 1495, v, 760, 745, 710, 700 cm.- in Nujol; NMR confirmed thestructure.

Analysis.-Calcd. for C H NO (percent): C, 77.88; H, 9.15; N, 6.05. Found(percent): C, 77.84; H, 9.45; N, 6.13.

Example 39.-3-benzyl-3-azabicyclo[3.2.2]nonanendo-6-ol hydrochloride The3-benzoyl 3 azabicyclo[3.2.2]nonan-endo-6-ol (6.69 g.) was dissolved in80 ml. of tetrahydrofuran and added with stirring to a mixture of 6.0 g.of lithium aluminum hydride in 100 ml. of ether. The mixture wasrefluxed for one hour, chilled in a cold bath, and carefully decomposedby the addition of 25 ml. of water. After dilution with 300 ml. of etherand filtering, the filtrate and ether wash was dried (MgSO and thesolvent removed under reduced pressure to give 5.90 g. of 3-benzyl-3azabicyclo[3.2.2]nonan-endo-6-ol as a straw colored oil. Part of theoil (1.33 g.) was dissolved in ether and treated with ethereal hydrogenchloride to precipitate the salt which was recrystallized from methanolether to give 1.07 g. of 3-benzyl-3-azabicyclo[3.2.2]nonan-endo-6-olhydrochloride, M.P. 185-187 C.

Analysis.-Calcd. for C H NOCl (percent): N, 5.23;

Cl, 13.24. Found (percent): N, 5.47; Cl, 13.74.

Example 40.3-benzyl-3-azabicyclo [3.3 .1 ]nonanaxial-6-ol A solution of3-benzoyl-3-azabicyclo[3.3.1]nonanaxial-6-ol (5.0 g., 0.0204 mole) intetrahydrofuran (100 ml.) was poured into a mixture of lithium aluminumhydride (3.0 g.) and tetrahydrofuran (150 ml.). The whole mixture washeated at reflux temperature for five hours then the excess hydride wasconsumed with ethyl acetate and water. The inorganic solids were removedby filtration and washed with hot tetrahydrofuran. The tetrahydrofuransolution was dried over magnesium sulfate and concentrated under reducedpressure to an oil. The oil was transferred with ether to a distillationflask. After a few minutes at low pressure, the oil solidified. Thesolid crystallized from cold hexane, giving crystals, M.P. 67-69 C.Three recrystallizations from cold hexane, the last preceded bydecolorization with activated charcoal, resulted in colorless crystalsof 3-benzyl-3-azabicy- 26 clo[3.3.1]nonan-axial-6-ol, M.P. 7071 0; v3320, v 1600, 1495, 11,, 730, 695 cm: on the oil.

Analysis.Calcd. for C H NO (percent): C, 77.88; H, 9.15; N, 6.05. Found(percent): C, 78.01; H, 9.54; N, 6.33.

Example 41 .-3-benzyl-3 -azabicyclo [3 .2.2] nonan- 6-one hydrochlorideThe 3-benzoyl-3-azabicyclo[3.2.2]nonan-6-one (5.13 g.), 60 ml. ofbenzene, 8 ml. of pyrrolidine, and mg. of p-toluenesulfonic acid washeated at reflux under a water trap for two hours. The mixture wasconcentrated under reduced pressure to a dark oil. A solution of thecrude enamine in 50 ml. of ether, and 25 ml. of tetrahydrofuran wasadded to a stirred mixture of 3.0 g. of lithium aluminum hydride and 100ml. of ether. After stirring at reflux for one hour the mixture waschilled and cautiously treated with 15.0 ml. of water, diluted withether, and filtered. The filtrate was dried (MgSO and concentrated toyield a light yellow oil. Infrared indicated that the enamine protectivemoiety has been replaced by carbonyl. The oil thus obtained wasdissolved in ether and treated with ethereal HCl to precipitate3-benzyl-3-azabicyclo[3.2.2]nonan-6-one hydrochloride.

Analysis.-Calcd. for C H N-0-HCl (percent): N, 5.27;

CI, 13.34. Found (percent): N, 5.57; Cl, 13.55.

Example 42.3-benzyl-3-azabicyclo [3.2.2] nonan-4-ol The crude3-benzyl-3-azabicyclo[3.2.2]nonan-4-ol resulting from the hydridereduction of 10 g. of 4-hydroxy- 3-benzoyl-3-azabicyclo[3.2.2]nonane,dissolved in 600 ml. of toluene and 150 ml. of cyclohexanone wasdistilled to remove ca. 100 ml. of toluene. Aluminum isopropoxide (20g.) was added, the mixture was distilled to remove ca. 50 m1. of solventand then heated at reflux for 60 minutes. After cooling it was pouredonto an ice mixture containing excess hydrochloric acid, stirred, andthe layers were separated. The aqueous acid layer was extracted severaltimes with ether and then made basic With 50% sodium hydroxide solution.The resulting emulsion was well extracted with ether and the etherextract was washed once with water and dried over magnesium sulfate. Theether solution was made up to 650 ml. and 100 ml. of this was treatedwith ethereal HCl to precipitate the salt, which was recrystallized frommethanol-ether to give 0.744 g. of3-benzyl-3-azabicyclo[3.2.2]nonan-6-one hydrochloride.

Analysis.Calcd. for C H NO-HCl (percent): C, 67.78; H, 7.59; N, 5.27;Cl, 13.34. Found (percent): C, 68.13; H, 7.77; N, 5.42; Cl, 13.17.

Example 43.3-benzyl-3-azabicyclo [3 .3 .1]nonan-6-one A solution of3-benzyl-3-azabicyclo[3.3.1]nonan-6-ol obtained by lithium aluminumhydride reduction of 7.457 g. of3-benzoyl-3-azabicyclo[3.3.1]nonan-6-one in toluene ml.) and cyclohexane(30 ml.) was heated to boiling and the toluene-water azeotrope distilledoff. Aluminum i-propoxide (10 g.) and cyclohexanone (10 ml.) were addedto the solution and the mixture was heated at reflux temperature for twohours. The mixture was poured into ice-aqueous hydrochloric acid andstirred. The aqueous layer was separated, extracted with ether (3X 100ml.), and made alkaline with concentrated sodium hydroxide solution. Aheavy precipitate formed at the neutralization point but disappeared andan oil formed as additional base was added. The solution and oily phasewere extracted (3X 100 ml.) with ether; the ether was dried overmagnesium sulfate and concentrated under reduced pressure to areddish-brown oil. The oil was transferred with ether to a 10 ml.distillation flask and distilled, M.P. 126-129 C. (0.04 mm.), giving4.171 g. (0.0182 mole, 60% yield) of 3-benzyl-3-azabicyclo[3.3.1]

nonan-6-one of colorless oil, n 1.5499; 11 1700, 11 1600, 1580, 1490, v,735, 695 cmr on the oil.

Analysis.-Calcd. for C H NO (percent): C, 78.56; H, 8.35; N, 6.11. Found(percent): C, 78.90; H, 8.56; N, 6.05.

Example 44.-3-benzyl-3-azabicyclo [3 .3 .1]nonan- 6-one perchlorate From3-benzyl-3 -azabicyclo[3.3.1jnonan-fi-one ethylene ketal.Aqueousperchloric acid (70%, 15 drops) was added to a solution of3-benzyl-3-azabicyclo[3.3.1]nonan- 6-one ethylene ketal (0.25 g.0.000915 mole) in absolute ethanol (5.0 ml.). The solution was heated onthe steam bath for three minutes. Addition of ether slowly precipitatedan oily solid, which crystallized into colorless crystals (0.251 g.,0.000763 mole, 83%), M.P. 210-215 C. Two recrystallizations from ethanolcontaining a few drops of water gave crystals of 3-benzyl-3-azabicyclo-[3.3.1]nonan-6-one perchlorate, M.P. 213-216 C., VNH +3080, v 1695, 111500, 1 770, 745, 705 cm.- in Nujol.

Analysis.Calcd. for C H NO Cl (percent): C, 54.63;

H, 6.11; N, 4.25. Found (percent): C, 54.51; H, 5.87;

From 3 benzyl 3 azabicyclo[3.3.1]nonan-6-one. Aqueous perchloric acid(70%, 10 drops) was added to a solution of3-benzyl-3-azabicyclo[3.3.1]nonan-6-one (0.236 g., 0.00103 mole) inabsolute ethanol (3 ml.). Crystals formed after 10 minutes. A first cropof 0.157 g. of colorless crystals was collected by filtration. A secondcrop of 0.042 g. (0.199 g. total, 0.000605 mole, 58%) was obtained fromthe mother liquor. The infrared spectrum of the crystals of3-benzyl-3-azabicyclo[3.3.1]nonan-6-one perchlorate thus obtained wasidentical to that of the above salt.

Example 45 .--3 -benzoyl-7-bromo-3 -azabicyclo 3 .3 1 nonan-6-one Asolution of bromine in chloroform was added in small portions to a cold(5l5 C.) solution of 3-benzoyl-3- azabicyclo[3.3.1]nonan-6-one (2.489g., 10.2 mmole) in chloroform (50 ml.) until the solution retained alight yellow color longer than 1-2 minutes. The solution was washed withwater, 5% aqueous sodium bicarbonate (remained basic to pH paper), andagain with water. The solution was dried (magnesium sulfate) andconcentrated to an oil. The oil was crystallized fromacetone-Skellysolve B, giving 2.381 g. (7.40 mmole, 72%) of product,M.P. 141-144 C. Two recrystallizations, the last preceded bydecolorization with activated charcoal, from acetone-Skellysolve B gavecolorless crystals of 3-benzoyl- 7-bromo-3-azabicyclo[3.3.1]nonan-6-one,M.P. 147-149 C.; 11 1715, 1630, v 1590, 1575, 1490, v, 730, 715, 700cm.- in Nujol.

Analysis.Calcd. for C 'H BrNO (percent): C, 55.91; 'H, 5.01; N, 4.35;Br, 24.80. Found (percent): C, 56.09; H, 5.28; N, 4.59; Br, 25.05.

Example 46.3 -benzoyl-7-methoxy-3-azabicyclo [3.3. 1 nonan-6-one Asolution of 3-benzoyl-7-bromo-3-azabicyclo[3.2.2] nonan-fi-one (1.888g., 5.86 mmoles) in methanol was mixed with a solution of sodiumhydroxide (0.485 g., 12.1 mmoles) in methanol (25 ml.). The solution waskept in the dark at room temperature for hours. The reaction solutionwas concentrated under reduced pressure to an oily solid, which waswashed with ethyl acetate. The ethyl acetate wash was placed on a columnof silica gel (150 g.) packed with ethyl acetate, and eluted with ethylacetate. Fractions 68 were combined in acetone. Addition of SkellysolveB resulted in formation of crystals over a period of about 20 hours. Thecrystals (0.142 g.) thus obtained were collected and recrystallized from28 acetone-Skellysolve B to give colorless crystals of 3-benzoyl-7-methoxy-3-azabicyc1o [3.3.1 nonan-6-one, M.P. 114-116" C.; v1710, 1625, 11 1600, 1490, u, 775, 745, 700 cm. in Nujol; NM'R confirmedthe assigned structure.

Analysis.-Calcd. for C H NO (percent): C, 70.31; H, 7.01; N, 5.13. Found(percent): C, 70.08; H, 7.25; N, 5.39.

Example 47.-3-benzoyl-6-(N-pyrrolidyl)-3-azabicyclo [3.2.2]nonane,hydrochloride A mixture of 3-benzoyl-3-azabicyclo[3.2.2]nonan-6-one (11g.) ml. of benzene, 16 ml. of pyrrolidine, and 200 mg. ofp-toluenesulfonic acid was stirred and heated at reflux under a watertrap for 3.5 hours and allowed to stand at 25 for 18 hours. The solventwas removed under reduced pressure on a hot water bath. The residualenamine was taken up in 100 ml. of ethanol and shaken with 1.0 g. of 10%palladium on carbon and hydrogen (40 p.s.i.g.) for 50 minutes. Thecatalyst was removed by filtration and the combined filtrate and washsolvent was concentrated under reduced pressure. The residue concentratethus obtained was taken up in 10% hydrochloric acid and extracted withether. The aqueous acid solution was made basic with 10% sodiumhydroxide solution and extracted with ether. This latter extract wasdried (Na SO and concentrated to give 14.6 g. of an oil. The oil waschromatographed over 500 g. of Florisil eluting with 7. 8 1. of solvent.Skellysolve B containing increasing proportions of acetone from 0 to30%, and collecting fractions of ml. each. Cuts 19-55 contained3-benzoyl-6- (N-pyrrolidyl) -3-azabicyclo [3 .2.2] nonane as determinedby TLC and IR; these were pooled in ether, dried over sodium sulfate andtreated with ethereal hydrogen chloride to precipitate the aminehydrochloride, which was recovered, washed with ether, and crystallizedfrom acetone to yield 5.72 g. of 3-benzoyl-6-(N- pyrrolidyl) 3azabicyclo[3.2.2]nonane, hydrochloride, M.P. 228-230 C.

Analysis.--Calcd. for C H N OCI (percent): C, 68.14; H, 8.13; N, 8.37;Cl, 10.59. Found (percent): C, 68.39; H, 8.38; N, 8.18; Cl, 10.37.

Example 48.6-(N-pyrrolidyl)-3-azabicyclo[3.2.2] nonane dimaleate The 3benzoyl '6 (N-pyrrolidyl)-3-azabicyclo[3.2.2] nonane obtained from 3.5g. of the corresponding hydrochloride salt obtained from Example 48,above, by treatment with aqueous sodium hydroxide and extraction withether and drying (Na SO was added to a stirred mixture of 3.0 g. ofLiAlH, in 100 ml. of ether. The mixture was heated at reflux for twohours, chilled in a cold bath, and decomposed by the addition of 20 ml.of water. After dilution with more ether the mixture was filtered, andthe filtrate and ether wash were dried ('MgS0 This solution was made upto 400 ml. with ether and 300 ml. of this solution ether was taken todryness, the free base thus obtained was dissolved in 50 ml. of ethanoland shaken with 0.5 g. of 10% Pd on C and hydrogen (40 p.s.i.g.) for 17hours. The catalyst was removed by filtration and the solvent removedunder reduced pressure to yield 6-(N-pyrrolidyl)-3-azabicyclo[3.2.2]nonane as an oil. This was made up to 100ml. with ether. Ten milliliters of this solution was used to precipitatethe HCl salt which proved to be very hygroscopic. Two milliliters ofether solution was used to prepare the p-toluenesulfonate by adding afew drops of ethanol solution of p-toluenesulfonic acid.

The remaining ether solution was treated with a methanolic solution ofmaleic acid to precipitate the maleate salt which was recovered andrecrystallized from methanol-methyl ethyl ketone to yield 1.54 g. of6-(N- pyrrolidyl)-3-azabicyclo[3.2.2]nonane dimaleate, 167168 C.

Example 49.6-methyl-3-azabicyclo[3.2.2]nonan-exo- 6-01, hydrochlorideTen grams of 3-benzoyl-3-azabicyclo[3.2.2]nonan-exo- 6-ol in 200 ml. oftetrahydrofuran was added to a stirred solution of 100 ml. of 3 Mmethylmagnesium bromide in ether. The mixture was distilled until thevapor temperature was 60 and then heated at reflux for 4.5 hours. Thestirred mixture was chilled and treated with 60 ml. of water, followedby 50 ml. of acetic acid and extracted several times with ether. Theaqueous solution from the ether extraction was made basic with 50%sodium hydroxide and the resulting gelatinous mixture was continuouslyextracted with ether, the extract dried (Na SO and treated with hydrogenchloride. The resulting HCl salt was recovered and washed with ether toyield 3.29 g. of 6-methyl-3-azabicyclo[3.2.2]nonan-exo-6-olhydrochloride, M.P. 228-230 C., which was recrystallized from methanolether, M.P. 230232 C.

Analysis.--Calcd. for C H NOCI (percent); C, 56.38; H, 9.46; N, 7.30;Cl, 18.50. Found (percent): C, 56.67; H, 9.97; N, 8.09; Cl, 18.63.

Example 50.-"-6-phenyl-3-azabicyclo[3.2.2]nonanexo-6-ol, hydrochloride Asolution of 20 g. of 3-benzoyl-3-azabicyclo[3.2.2] nonan-exo-6-ol in 400ml. of tetrahydrofuran was added to a stirred solution of 200 ml. of 3 Mphenylmagnesium' bromide in ether. Solvent was removed by distillationuntil the boiling temperature was 60 and the mixture was heated atreflux for 4.5 hours. After cooling, it was poured onto ice and stirredand acidified with concentrated hydrochloride while continuing to addice. This mixture was extracted several times with ether. The aqueousacid solution was made basic with 50% sodium hydroxide solution and theresulting mixture was extracted several times with ether. The etherextract was dried (MgSO treated with ethereal hydrogen chloride, and theinsoluble salt was recovered and washed with ether to yield 15.32 g.This was fractionally crystallized from methanol-methyl ethyl ketone togive about 7.5 g. of 6-phenyl-3-azabicyclo- E3.2.2]nonan-exo-6-olhydrochloride, M.P. 238-240 C.

dec.).

Analysis.--Calcd. for C H NOCI (percent): C, 66.26; H, 7.94; N, 5.52;Cl, 13.97. Found (percent): C, 66.51; H, 8.17; N, 5.49; Cl, 13.90.

The filtrates from above produced about 2.0 g. of 6- phenyl 3azabicyclo[3.2.2]nonan-endo-6-ol hydrochloride, M.P. 218220 C. (dec.).

Analysis.Calcd. for C H NOCl (percent): C, 66.26; H, 7.94; N, 5.52; Cl,13.97. Found (percent): C, 66.09; H, 8.33; N, 5.36; Cl, 13.70.

Example 51.--1,4-dioxa-8-azaspiro[4.6]undecane A solution of 8 benzyl1,4-dioxa8-azaspiro[4.6] undecane (8.692 g., 0.0352 mole) in methanol(130 ml.) was shaken with hydrogen and palladium-on-carbon (2.50 g.)until the uptake of hydrogen ceased. The catalyst was removed byfiltration and washed with methanol. The combined methanol solution wasconcentrated under reduced pressure to an oil. Simple distillation ofthe oil gave 5.047 g. (0.032 mole, 91% yield) of 1,4-dioxa-8-azaspiro[4.6]undecane as a colorless oil, B.P. 58-60 C. (0.05 mm.); r21.4885; 1 3330 cm.- neat.

Analysis.Calcd. for C H NO (157.21) (percent): C, 61.12; H, 9.62; N,8.91. Found (percent): C, 61.17; H, 9.86; N, 8.85.

Example 52.8-p-toluenesulfonyl-1,4-dioxa-8-azaspiro- [4.6]undecane Asolution of 1,4-dioxa-8-azaspiro[4.6]undecane (0.622 g., 0.00396 mole)in 16% aqueous sodium hydroxide was mixed with p-toluenesulfonylchloride (0.786 g., 0.00413 mole). The mixture was shaken vigorously forseveral minutes, then warmed and shaken more. The mixture was left atroom temperature overnight and then was extracted with ether (2X 20ml.). The ether solution was dried and concentrated to give8-p-toluenesulfonyl-1,4- dioxa-8-azaspiro[4.6]undecane as an oil.

Example 53.1,4-dioxa-9-azaspiro[4.7] dodecane A solution of 9 benzyl1,4-dioxa-9-azaspiro[4.7] dodecane (24.642 g., 0.0945 mole) in absoluteethanol (150 ml.) was shaken with 5% palladium-on-carbon (5.88 g.) andhydrogen in a hydrogenation apparatus. After 30 minutes the hydrogenuptake had stopped and totaled 28 pounds (calculated, 27.4 pounds). Thecatalyst was removed by filtration. The catalyst was washed twice withethanol. Ethanol was removed .from the combined solvent and washed bydistillation. The residual oil was purified by a simple distillationwhich gave 1,4-dioxa- 9-azaspiro[4.7]dodecane as a colorless oil (14.447g., 0.0844 mole, 89%), B.P. 70-75 C. (0.2 mm.); n' 1.4835; 11 3360 cm.-on the oil.

Analysis.Calcd. for C H NO (171.23) (percent): C, 63.13; H, 10.00; N,8.18. Found (percent): C, 63.31; H, 10.02; N, 8.17.

Example 54.1,4-dioxa-8-azaspiro[4.7]dodecane A solution of 8 benzyl1,4-dioxa-8-azaspiro[4.7] dodecane (5.152 g., 0.0197 mole) in absoluteethanol ml.) was shaken with 5% palladium-on-carbon (1.5 g.) andhydrogen in a hydrogenation apparatus for 3 hours. Hydrogen uptake wascomplete after one hour. The catalyst was removed by filtration and waswashed twice with methanol. The excess solvent was removed bydistillation under reduced pressure. Distillation of the residual oilgave 2.771 g. (0.0162 mole, 82% yield) of 1,4-dioxa-8-azaspiro[4.7]dodecane as a colorless oil, B.P. 68-69" C. (0.1 mm.); n1.4853; u 3360 m.w. cmr on the oil.

Analysis.0alcd. for C H NO (171.23) (percent): C, 63.13; H, 10.00; N,8.18. Found (percent): C, 63.16; H, 10.04; N, 7.88.

Example 55.1,4-dioxa-9-azaspiro[4.8]tridecane A solution of 9 benzyl 1,4dioxa-9-azaspiro [4.8] tridecane (9.451 g., 0.0344 mole) in methanolml.) was shaken with 5% palladium-on-carbon (2.5 g.) in hydrogen for 45minutes at which time uptake of hydrogen appeared complete. The catalystwas removed by filtration and the colorless filtrate stored overnight inthe refrigerator. The solution was concentrated under reduced pressure.The oil thus obtained crystallized as it cooled and the solid wasdissolved in ether, decolorized with activated charcoal, filtered andcrystallized by addition of hexanes to the ether and by cooling in the.freezer to give 4.649 g. of 1,4 dioxa 9 azaspiro[4.8]tridecane (0.0251mole, 73% yield), M.P. 55-57 C.; recrystallization from etherhexanesgave colorless, chunky crystals of 1,4-dioxa-9- azaspiro[4.8]tridecane,M.P. 55-57 C.; r 3400 m.w. cm.- in Nujol.

Analysis.Calcd. for C H NO (185.26) (percent): C, 64.83; H, 10.34; N,7.56. Found (percent): C, 64.66; H, 10.40; N, 7.62.

Example 56.--3-azabicyclo[3.2.2]nonan-6-one hydrochloride 3 benzyl 3azabicyclo [3.2.2]nonan-6-one (4.55 g.) was dissolved in 90 ml. ofethanol and shaken with 1.0 g. of 10% palladium-on-carbon and hydrogen(50 p.s.i.g.) for minutes. The catalyst was removed by filtration;

the filtrate and wash were concentrated in vacuo to a small volume,diluted with ether, and treated with etheral HCl. The hydrochloride ofthe product was recovered, washed with ether and dried; yield, 3.12 g.of 3-azabicyclo[3.2.2] nonan-6-one hydrochloride, M.P. 218-220 C.(dec.); a sample from methanol-ether melted at 227-229 C. (dec.).

Analysis.-Calcd. for C H N-HC1 (percent): C, 54.70; H, 8.03; N, 7.98;Cl, 20.19. Found (percent): C, 54.22; H, 8.14; N, 7.98; Cl, 20.64.

Example 57.3-azabicyclo[3.2.2] nonan-endo-6-o1 hydrochloride A solutionof 9.17 g. of 3-benzyl-3-azabicyclo[3.2.2] nonan-endo-G-ol in 120 ml. ofethanol was shaken with 1.0 g. of 10% palladium carbon and hydrogen (44p.s.i.g. starting pressure for 20 hours. The mixture, freed of catalystand concentrated in vacuo gave 5.60 g. of3-azabicyclo[3.2.2]nonan-endo-6-ol. A portion of the free base wasdissolved in ether and treated with ethereal HCl to precipitate theamine hydrochloride, which was recrystallized from methanol-methyl ethylketone to give 3-azabicyclo[3.2.21nonan-endo-6-ol hydrochloride, M.P.280 C. (dec.).

Analysis.-Calcd. for C H NOCl (percent): C, 54.07; H, 9.08; N, 7.88; Cl,19.96. Found (percent): C, 54.17; H, 9.05; N, 8.01; Cl, 19.99.

Example 58.1,4-dioxa-9-azaspiro[4.7]dodec-9-yl acetonitrile A solutionof 1,4 dioxa 9 azaspiro[4.7]dodecane (10.215 g., 0.0597 mole) in benzene(25 ml.) was added slowly to a stirred mixture of chloroacetonitrile(6.0 g., 0.0795 mole) in benzene (125 ml.) and anhydrous sodiumcarbonate (4.0 g.). During the course of addition (15 minutes), themixture was warmed to near the reflux temperature and then was heated toreflux with stirring for 18 hours. The precipitate thus obtained wasdissolved in dilute aqueous sodium bicarbonate solution. The benzenelayer was washed twice with water and dried over magnesium sulfate.Concentration of the benzene under reduced pressure gave an oil whichcrystallized upon cooling. The crystalline material dissolved in hotSkellysolve B leading a small amount of gummy, yellow residue and acolorless solution. Cooling gave colorless crystals (8.960 g.), M.P.77-78 C. A second crop, M.P. 7S77 C., (1.726 g., total 10.6 86 g.,0.0508 mole, 85% yield) was obtained from the concentrated filtrate.Recrystallization from Skellysolve B gave l,4-dioxo-9-azaspiro[4.7]dodec- 9-yl acetonitrile as colorless needles, M.P. 78-79C.;

70 2220 cmr in Nujol.

| Example 59.-9-(2-aminoethyl)-1,4-dioxa-9-azaspiro [4.7]dodecane Asolution of :1,4-dioxa-9-azaspiro[4.7 dodec-9-y1 acetonitrile (8.095 g.,0.0385 mole) in ether (100 ml.) was added dropwise to a mixture oflithium aluminum hydride (5.0 g., 0.13 mole) in ether (200 ml). Themixture was heated to reflux for three hours with stirring and then wasstirred at room temperature for 16 hours. The excess lithium aluminumhydride was decomposed with acetonewater and the inorganic salts removedby filtration. The solids were washed by warming with ether (300 ml.).The combined ether filtrates were dried over magnesium sulfate andconcentrated to an oil which was distilled, giving 7.074 g. (0.0330mole, 86% yield) of 9-(2-amino- 32 ethyl)1,4-dioxa9-azaspiro[4.7]dodecane as a colorless oil, B.P. 95- 101" C.(0.10 mm.); u 1.4940; 1 3360, 3280 cm.- on the oil.

Analysis.-Calcd. for CHI-122N202 (214.30) (percent): C, 61.65; H, 10.35;N, 13.07. Found (percent): C, 61,28; H, 10.48; N, 13.22.

Example 60.--1-(-guanidinylethyl)-5-oxohetpamethyleneimine sulfateConcentrated sulfuric acid (1.17 g., 0.0120 mole) was added to asolution of 9-(Z-aminoethyl)-1,4-dioxaspiro- [4.7]dodecane (5.116 g.,0.0239 mole) in water (20 m1.). To this solution,2-methyl-2-thiopseudourea sulfate (3.34 g., 0.0120 mole) was added andthe resulting solution heated to reflux for 16 hours. Addition ofethanol to the cooled solution caused the slow formation of colorlesscrystals, first crop 4.129 g.; two additional crops formed upon additionof more ethanol, giving 1.595 g. (total 5.724 g., 0.0184 mole, 77%yield) of colorless crystals. The crops were combined and recrystallizedtwice from water-ethanol to give colorless crystals of1-(-guanidinylethyl)-5-oxoheptamethyleneimine sulfate, M.P. turningbrown to 300 G;

mm 3200 m.s, 3070 s., 2640 m.w.,

IIC=N 1113., 1635 LIL, 1600 m.w., cm.- in Nujol.

Analysis.-Calcd. for C H N O5S (310.38) (percent): C, 38.69; H, 7.15; N,18.05; S, 10.33. Found (percent): C, 38.82; H, 7.32; N, 18.07; S, 10.27.

Example '61 .3-cyc1ohexyl-3 -'azabicyclo[ 3 .2.2.] nonanendo-6-olhydrochloride A mixture of 3.40 g. of 3-azabicyclo[3.2.2]nonan-endo-6-ol, 50 ml. of benzene, 3.0 ml. of cyclohexanone, and mg. ofp-toluenesu1fonic acid was stirred and heated at reflux for 2.5 hours.The mixture was concentrated to dryness under reduced pressure and theresidual oil was taken up in Skellysolve B hexanes and filtered toremove insoluble material. The filtrate was concentrated under reducedpressure to an oil residue of enamine still containing cyclohexanone.This oil in 50 ml. of ethanol was shaken with 1.0 g. of 10% Pd on C andhydrogen (39 p.s.i.g.) for two hours. The catalyst was removed byfiltration and the filtrate concentrated under reduced pressure to givea crystalline residue of 3-cyclohexyl-3-azabicyclo[3.2.2]endo-6-ol whichwas dissolved in ether and treated with ethereal hydrogen chloride; theresulting HCl salt was recovered and recrystallized from methanolmethylethyl ketone; yield, 3.43 g. of3-cyclohexyl-3-azabicyclo[3.2.2]nonan-endo-6-ol, M.P. 265 C. (dec.).

Analysis.Calcd. for C H NOCI (percent): C, 64.71; H, 10.01; N, 5.39; Cl,13.65. Found (percent): C, 64.60; H, 10.21; N, 5.39; Cl, 13.74.

We claim: 1. 1-(Z-guanidinoethyl)-5-oxoheptamethyleneimine.

References Cited UNITED STATES PATENTS 2,775,589 12/ 1956 Diamond et a1.260-239 BF ALTON D. ROLLINS, Primary Examiner US. Cl. X.R.

